Hck as a therapeutic target in myd88 mutated diseases

ABSTRACT

The present disclosure provides compounds of Formula (I), and pharmaceutically acceptable salts, solvates, hydrates, polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeled derivatives, and prodrugs thereof. The provided compounds may be kinase (e.g., HCK, BTK, LYN, BLK, FRK) inhibitors. Also provided are pharmaceutical compositions and kits including the provided compounds. Further provided are methods of using the provided compounds, pharmaceutical compositions, and kits (e.g., for treating diseases (e.g., proliferative diseases) in a subject in need thereof).

RELATED APPLICATIONS

The present application claims priority under 35 U.S.C. § 119(e) to U.S.provisional application, U.S. Ser. No. 62/912,555, filed Oct. 8, 2019,which is incorporated herein by reference.

BACKGROUND

It has been discovered that Hematopoietic cell kinase (HCK)transcription and activation is triggered by mutated myeloiddifferentiation primary response 88 (MYD88), and is an importantdeterminant of pro-survival signaling. It has also been discovered thatinhibition of the kinase activity of HCK triggers apoptosis in mutatedMYD88 cells. For example, the expression of MYD88 mutations inWaldenström's Macroglobulinemia (WM), wherein 95-97% of patients expressMYD88^(L265P), and more rarely non-L265P MYD88 mutations. WM isconsidered to correspond to lymphoplasmacytic lymphoma (LPL) as definedby the World Health Organization classification system. Up to 30% ofpatients with Activated B-Cell (ABC) Subtype of Diffuse Large B-celllymphoma (ABC DLBCL) also express activating MYD88 mutations, includingMYD88^(L265P). Mutations in MYD88 promote Myddosome self-assembly andcan trigger NF-kB signaling in the absence of Toll (TLR) or IL1 (IL1R)receptor signaling through IL1 Receptor Associated Kinases (IRAK4/IRAK1)or Bruton's Tyrosine Kinase (BTK).

Next generation sequencing has revealed activating myeloiddifferentiation primary response 88 (MYD88) mutations in several B-cellmalignancies including Waldenström's macroglobulinemia (immunoglobulin M(IgM) secreting lymphoplasmacytic lymphoma), non-IgM secretinglymphoplasmacytic lymphoma, ABC subtype of diffuse large B-celllymphoma, primary central nervous system (CNS) lymphoma, immuneprivileged lymphomas that include testicular lymphoma, marginal zonelymphoma, and chronic lymphocytic leukemia. Particularly striking hasbeen the expression of MYD88 mutations in Waldenström'smacroglobulinemia (WM), wherein 95-97% of patients express MYD88^(L265P)and more rarely non-L265P MYD88 mutations. Waldenström'smacroglobulinemia is considered to correspond to lymphoplasmacyticlymphoma (LPL) as defined by the World Health Organizationclassification system. Up to 30% of patients with Activated B-Cell (ABC)Subtype of Diffuse Large B-cell lymphoma (ABC-DLBCL) also expressactivating MYD88 mutations, including MYD88^(L265P). Mutations in MYD88promote Myddosome self-assembly and can trigger NF-kB signaling in theabsence of Toll (TLR) or IL1 (IL1R) receptor signaling through IL1Receptor Associated Kinases (IRAK4/IRAK1) or Bruton's Tyrosine Kinase(BTK).

Ibrutinib is an inhibitor of BTK that is highly active in WM, resultingin responses in 91% of previously treated patients. In WM patients, bothmajor and overall responses to ibrutinib are higher in patients withMYD88 mutations. Ibrutinib also shows activity in previously treatedpatients with ABC DLBCL, particularly among patients with MYD88mutations. Ibrutinib is also active in other B-cell malignanciesincluding Chronic Lymphocytic Leukemia (CLL) and Mantle Cell Lymphoma(MCL). Suppression of tonic B-cell receptor (BCR) activity mediated byBTK has been implicated as the mechanism underlying ibrutinib activityin non-WM B-cell diseases.

SUMMARY OF THE INVENTION

Provided herein are compounds of Formula (I):

wherein J, A, R, R¹, R², and R³ are as defined herein, andpharmaceutically acceptable salts, solvates, hydrates, polymorphs,co-crystals, tautomers, stereoisomers, isotopically labeled derivatives,and prodrugs thereof.

In certain embodiments, a compound of Formula (I) is of formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

The provided compounds may be kinase (e.g., an SRC family kinase (i.e.SFK) (e.g., HCK, LYN, BLK, FRK), a Tec family kinase (e.g., BTK)) and incertain aspects, the compounds may be specific or selective for SFKs(e.g., HCK, LYN, BLK, FRK) or Tec family kinases (e.g., BTK) over one ormore other kinases. Also provided are pharmaceutical compositions andkits comprising the disclosed compounds. The present disclosure alsoprovides methods of using the disclosed compounds, pharmaceuticalcompositions, and kits (e.g., for treating a disease (e.g., aproliferative disease (e.g., an IgM gammopathy (e.g., an IgM Monoclonalgammopathy of undetermined significance (MGUS), amyloid light chain (AL)amyloidosis), mastocytosis (e.g., systemic mastocytosis), cancer (e.g.,breast cancer, colon cancer, testicular cancer, CNS cancer, stomachcancer, lymphoma (e.g., B-cell lymphoma (e.g., lymphoplasmacyticlymphoma (e.g., IgM secreting lymphoplasmacytic lymphoma (i.e.,Waldenström's Macroglobulinemia), non-IgM secreting lymphoplasmacyticlymphoma)), diffuse large B-cell lymphoma (e.g., activated B-cell-like(ABC)-DLBCL, germinal center B-cell-like (GBC)-DLBCL), follicularlymphoma, marginal zone B-cell lymphoma, small lymphocytic lymphoma,mantle cell lymphoma), myeloma (e.g., IgM myelomas (e.g., IgM multiplemyeloma)), and leukemia (e.g., chronic lymphocytic leukemia (CLL), acutelymphoblastic leukemia, myelogenous leukemia (e.g., chronic myelogenousleukemia, acute myelogenous leukemia (e.g., mast cell leukemia),myeloproliferative diseases (e.g., myelodysplastic syndrome))))) in asubject in need thereof, or inhibiting the activity of a kinase in asubject in need thereof, in a biological sample, or in a cell).

In yet another aspect, the present disclosure provides compounds ofFormula (I), and pharmaceutically acceptable salts, solvates, hydrates,polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeledderivatives, and prodrugs thereof, and pharmaceutical compositionsthereof, for use in the treatment and/or prevention of a disease (e.g.,a proliferative disease, such as an IgM gammopathy, mastocytosis, orcancer) in a subject in need thereof.

In another aspect, the present disclosure provides uses of compounds ofFormula (I), and pharmaceutically acceptable salts, solvates, hydrates,polymorphs, co-crystals, tautomers, stereoisomers, isotopically labeledderivatives, and prodrugs thereof, and pharmaceutical compositionsthereof, in the manufacture of a medicament for treating and/orpreventing a disease in a subject in need thereof.

In another aspect, the present disclosure provides methods of preparinga compound of Formula (I), or a pharmaceutically acceptable salt,solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer,isotopically labeled derivative, or prodrug thereof.

In one aspect, the present disclosure provides pharmaceuticalcompositions including a compound described herein, and optionally apharmaceutically acceptable excipient. In certain embodiments, thepharmaceutical composition further comprises an additionalpharmaceutical agent. In certain embodiments, the additionalpharmaceutical agent is selected from the group consisting ofchemotherapy drugs, epigenetic modifiers, glucocorticoids, biologics,and immunotherapy agents. In another aspect, the additionalpharmaceutical agents is a BCL-2 inhibitor (e.g., venetoclax,navitoclax, obatoclax).

The pharmaceutical compositions may be useful for treating a disease ina subject in need thereof, inhibiting the activity of a kinase in asubject in need thereof, a biological sample, or a cell. In certainaspects, the disease is a proliferative disease (e.g., an IgM gammopathy(e.g., an IgM Monoclonal gammopathy of undetermined significance (MGUS),amyloid light chain (AL) amyloidosis), mastocytosis (e.g., systemicmastocytosis), cancer (e.g., breast cancer, colon cancer, testicularcancer, CNS cancer, stomach cancer, lymphoma (e.g., B-cell lymphoma(e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacyticlymphoma (i.e., Waldenström's Macroglobulinemia), non-IgM secretinglymphoplasmacytic lymphoma)), diffuse large B-cell lymphoma (e.g.,activated B-cell-like (ABC)-DLBCL, germinal center B-cell-like(GBC)-DLBCL), follicular lymphoma, marginal zone B-cell lymphoma, smalllymphocytic lymphoma, mantle cell lymphoma), myeloma (e.g., IgM myelomas(e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocyticleukemia (CLL), acute lymphoblastic leukemia, myelogenous leukemia(e.g., chronic myelogenous leukemia, acute myelogenous leukemia (e.g.,mast cell leukemia), myeloproliferative diseases (e.g., myelodysplasticsyndrome))))).

Provided herein are methods of treating a disease a proliferativedisease (e.g., an IgM gammopathy (e.g., an IgM Monoclonal gammopathy ofundetermined significance (MGUS), amyloid light chain (AL) amyloidosis),mastocytosis (e.g., systemic mastocytosis), cancer (e.g., breast cancer,colon cancer, testicular cancer, CNS cancer, stomach cancer, lymphoma(e.g., B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgMsecreting lymphoplasmacytic lymphoma (i.e., Waldenström'sMacroglobulinemia), non-IgM secreting lymphoplasmacytic lymphoma)),diffuse large B-cell lymphoma (e.g., activated B-cell-like (ABC)-DLBCL,germinal center B-cell-like (GBC)-DLBCL), follicular lymphoma, marginalzone B-cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma),myeloma (e.g., IgM myelomas (e.g., IgM multiple myeloma)), and leukemia(e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia,myelogenous leukemia (e.g., chronic myelogenous leukemia, acutemyelogenous leukemia (e.g., mast cell leukemia), myeloproliferativediseases (e.g., myelodysplastic syndrome))))) in a subject in needthereof, comprising administering to the subject a therapeuticallyeffective amount of a compound of Formula (I). In certain aspects,provided herein are methods for inhibiting a mutated BTK (e.g., a C481Smutant) in a subject in need thereof.

The present disclosure provides methods of treating a disease in subjectby administering to a subject in need thereof an effective amount of acompound, or a pharmaceutical composition thereof, as described herein.In certain aspects, the disease is a proliferative disease (e.g., an IgMgammopathy (e.g., an IgM Monoclonal gammopathy of undeterminedsignificance (MGUS), amyloid light chain (AL) amyloidosis), mastocytosis(e.g., systemic mastocytosis), cancer (e.g., breast cancer, coloncancer, testicular cancer, CNS cancer, stomach cancer, lymphoma (e.g.,B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secretinglymphoplasmacytic lymphoma (i.e., Waldenström's Macroglobulinemia),non-IgM secreting lymphoplasmacytic lymphoma)), diffuse large B-celllymphoma (e.g., activated B-cell-like (ABC)-DLBCL, germinal centerB-cell-like (GBC)-DLBCL), follicular lymphoma, marginal zone B-celllymphoma, small lymphocytic lymphoma, mantle cell lymphoma), myeloma(e.g., IgM myelomas (e.g., IgM multiple myeloma)), and leukemia (e.g.,chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia,myelogenous leukemia (e.g., chronic myelogenous leukemia, acutemyelogenous leukemia (e.g., mast cell leukemia), myeloproliferativediseases (e.g., myelodysplastic syndrome))))). Also described aremethods for contacting a biological sample or cell with an effectiveamount of a compound, or pharmaceutical composition thereof, asdescribed herein.

In certain embodiments, a method described herein further includesadministering to the subject in need thereof an additionalpharmaceutical agent. In certain embodiments, a method described hereinfurther includes contacting the biological sample or cell with anadditional pharmaceutical agent.

In one aspect, provided are methods for treating a disease in a subjectwho is resistant to treatment with a BTK inhibitor (e.g., ibrutinib,CC-292, ONO-4059, evobrutinib, spebrutinib, BGB-3111, HM71224, orACP-196 (i.e., acalabrutinib)). In certain aspects, a subject who isresistant to treatment with a BTK inhibitor has a mutated BTK (e.g., aC481S mutated BTK). In another aspect, the subject who is resistant totreatment with a BTK inhibitor is diagnosed to have an MYD88 mutateddisease (e.g., a proliferative disease (e.g., IgM gammopathy,mastocytosis, cancer)).

In another aspect, provided herein are methods of inhibiting a kinase(e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinases (e.g., BTK))in a subject in need thereof, comprising administering to the subject atherapeutically effective amount of a compound of Formula (I), or apharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, isotopically-labeled derivative, stereoisomer, or prodrugthereof. In certain aspects, provided herein are methods for inhibitinga mutated BTK (e.g., a C481S mutant) in a subject in need thereof.

In certain aspects, the method further comprises administering ananti-cancer agent to the subject. In some embodiments, the anti-canceragent is a chemotherapeutic agent. In another aspect, the method furthercomprises administering to the subject one or more of a proteasomeinhibitor (e.g., bortezomib, carfilzomib, ixazomib or oprozomib), amonoclonal antibody (e.g., rituximab, daratumumab, ofatumumab orobinituzumab), an alkylating drug (e.g., bendamustine,cyclophosphamide), a nucleoside analogue (e.g., fludarabine orcladribine), an mTOR inhibitor (e.g., everolimus), a BTK inhibitor(e.g., ibrutinib, acalabrutinib or BGB-3111), a BCR inhibitor (e.g., aSYK inhibitor), and/or an immunomodulatory agent (e.g., thalidomide orlenalidomide).

In another aspect, the present disclosure provides kits comprising: acompound of Formula (I), or a pharmaceutically acceptable salt, solvate,hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopicallylabeled derivative, or prodrug thereof, or a pharmaceutical compositionthereof; and instructions for using the compound, or a pharmaceuticallyacceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer,stereoisomer, isotopically labeled derivative, or prodrug thereof, orthe pharmaceutical composition.

The details of one or more embodiments of the present disclosure are setforth herein. Other features, objects, and advantages of the presentdisclosure will be apparent from the Detailed Description, Examples,Figures, and Claims.

Definitions

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Michael B. Smith, March's Advanced Organic Chemistry, 7^(th) Edition,John Wiley & Sons, Inc., New York, 2013; Richard C. Larock,Comprehensive Organic Transformations, John Wiley & Sons, Inc., NewYork, 2018; and Carruthers, Some Modern Methods of Organic Synthesis,3^(rd) Edition, Cambridge University Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers,and thus can exist in various stereoisomeric forms, e.g., enantiomersand/or diastereomers. For example, the compounds described herein can bein the form of an individual enantiomer, diastereomer or geometricisomer, or can be in the form of a mixture of stereoisomers, includingracemic mixtures and mixtures enriched in one or more stereoisomer.Isomers can be isolated from mixtures by methods known to those skilledin the art, including chiral high pressure liquid chromatography (HPLC),supercritical fluid chromatography (SFC), and the formation andcrystallization of chiral salts; or preferred isomers can be prepared byasymmetric syntheses. See, for example, Jacques et al., Enantiomers,Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen etal., Tetrahedron 33:2725 (1977); Eliel, E. L., Stereochemistry of CarbonCompounds (McGraw-Hill, N Y, 1962); and Wilen, S. H., Tables ofResolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ.of Notre Dame Press, Notre Dame, Ind. 1972). The present disclosureadditionally encompasses compounds described herein as individualisomers substantially free of other isomers, and alternatively, asmixtures of various isomers.

In a formula, the bond

is a single bond, the dashed line

is a single bond or absent, and the bond

or

is a single or double bond.

Unless otherwise provided, a formula depicted herein includes compoundsthat do not include isotopically enriched atoms and also compounds thatinclude isotopically enriched atoms. Compounds that include isotopicallyenriched atoms may be useful as, for example, analytical tools, and/orprobes in biological assays.

The term “aliphatic” includes both saturated and unsaturated,nonaromatic, straight chain (i.e., unbranched), branched, acyclic, andcyclic (i.e., carbocyclic) hydrocarbons. In some embodiments, analiphatic group is optionally substituted with one or more functionalgroups (e.g., halo, such as fluorine). As will be appreciated by one ofordinary skill in the art, “aliphatic” is intended herein to includealkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynylmoieties.

When a range of values (“range”) is listed, it is intended to encompasseach value and sub-range within the range. A range is inclusive of thevalues at the two ends of the range unless otherwise provided. Forexample, “an integer between 1 and 4” refers to 1, 2, 3, and 4. Forexample “C₁₋₆ alkyl” is intended to encompass, C₁, C₂, C₃, C₄, C₅, C₆,C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆, C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄,C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

A “hydrocarbon chain” refers to a substituted or unsubstituted divalentalkyl, alkenyl, or alkynyl group. A hydrocarbon chain includes (1) oneor more chains of carbon atoms immediately between the two radicals ofthe hydrocarbon chain; (2) optionally one or more hydrogen atoms on thechain(s) of carbon atoms; and (3) optionally one or more substituents(“non-chain substituents,” which are not hydrogen) on the chain(s) ofcarbon atoms. A chain of carbon atoms consists of consecutivelyconnected carbon atoms (“chain atoms”) and does not include hydrogenatoms or heteroatoms. However, a non-chain substituent of a hydrocarbonchain may include any atoms, including hydrogen atoms, carbon atoms, andheteroatoms. For example, hydrocarbon chain —C^(A)H(C^(B)H₂C^(C)H₃)—includes one chain atom C^(A), one hydrogen atom on C^(A), and non-chainsubstituent —(C^(B)H₂C^(C)H₃). The term “C_(x) hydrocarbon chain,”wherein x is a positive integer, refers to a hydrocarbon chain thatincludes x number of chain atom(s) between the two radicals of thehydrocarbon chain. If there is more than one possible value of x, thesmallest possible value of x is used for the definition of thehydrocarbon chain. For example, —CH(C₂H₅)— is a C₁ hydrocarbon chain,

and is a C₃ hydrocarbon chain. When a range of values is used, themeaning of the range is as described herein. For example, a C₃₋₁₀hydrocarbon chain refers to a hydrocarbon chain where the number ofchain atoms of the shortest chain of carbon atoms immediately betweenthe two radicals of the hydrocarbon chain is 3, 4, 5, 6, 7, 8, 9, or 10.A hydrocarbon chain may be saturated (e.g., —C≡C— or —(CH₂)₄—). Ahydrocarbon chain may also be unsaturated and include one or more C═Cand/or C≡C bonds anywhere in the hydrocarbon chain. For instance,—CH═CH—(CH₂)₂—, —CH₂—C≡C—CH₂—, and —C≡C—CH═CH— are all examples of aunsubstituted and unsaturated hydrocarbon chain. In certain embodiments,the hydrocarbon chain is unsubstituted (e.g., —(CH₂)₄—). In certainembodiments, the hydrocarbon chain is substituted (e.g., —CH(C₂H₅)— and—CF₂—). Any two substituents on the hydrocarbon chain may be joined toform an optionally substituted carbocyclyl, optionally substitutedheterocyclyl, optionally substituted aryl, or optionally substitutedheteroaryl ring. For instance,

are all examples of a hydrocarbon chain. In contrast, in certainembodiments

are not within the scope of the hydrocarbon chains described herein.When a chain atom of a C_(x) hydrocarbon chain is replaced with aheteroatom, the resulting group is referred to as a C_(x) hydrocarbonchain wherein a chain atom is replaced with a heteroatom, as opposed toa C_(x-1) hydrocarbon chain.

For example,

is a C₃ hydrocarbon chain, wherein one chain atom is replaced with anoxygen atom.

The term “alkyl” refers to a radical of a straight-chain or branchedsaturated hydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀alkyl”). In some embodiments, an alkyl group has 1 to 12 carbon atoms(“C₁₋₁₂ alkyl”). In some embodiments, an alkyl group has 1 to 10 carbonatoms (“C₁₋₁₀ alkyl”). In some embodiments, an alkyl group has 1 to 9carbon atoms (“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1to 8 carbon atoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl grouphas 1 to 7 carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkylgroup has 1 to 6 carbon atoms (“C₁₋₆ alkyl”). In some embodiments, analkyl group has 1 to 5 carbon atoms (“C₁₋₅ alkyl”). In some embodiments,an alkyl group has 1 to 4 carbon atoms (“C₁₋₄ alkyl”). In someembodiments, an alkyl group has 1 to 3 carbon atoms (“C₁₋₃ alkyl”). Insome embodiments, an alkyl group has 1 to 2 carbon atoms (“C₁₋₂ alkyl”).In some embodiments, an alkyl group has 1 carbon atom (“C₁ alkyl”). Insome embodiments, an alkyl group has 2 to 6 carbon atoms (“C₂₋₆ alkyl”).Examples of C₁₋₆ alkyl groups include methyl (C₁), ethyl (C₂), n-propyl(C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄),iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl(C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅, and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈)and the like. Unless otherwise specified, each instance of an alkylgroup is independently optionally substituted, e.g., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents. In certain embodiments, the alkyl group isunsubstituted C₁₋₁₂ alkyl (e.g., —CH₃ (Me), unsubstituted ethyl (Et),unsubstituted propyl (Pr, e.g., unsubstituted n-propyl (n-Pr),unsubstituted isopropyl (i-Pr)), unsubstituted butyl (Bu, e.g.,unsubstituted n-butyl (n-Bu), unsubstituted tert-butyl (tert-Bu ort-Bu), unsubstituted sec-butyl (sec-Bu or s-Bu), unsubstituted isobutyl(i-Bu)). In certain embodiments, the alkyl group is substituted C₁₋₁₂alkyl (such as substituted C₁₋₆ alkyl, e.g., —CH₂F, —CHF₂, —CF₃,—CH₂CH₂F, —CH₂CHF₂, —CH₂CF₃, or benzyl (Bn)). The attachment point ofalkyl may be a single bond (e.g., as in —CH₃), double bond (e.g., as in═CH₂), or triple bond (e.g., as in ═CH). The moieties=CH₂ and ≡CH arealso alkyl.

In some embodiments, an alkyl group is substituted with one or morehalogens. “Perhaloalkyl” is a substituted alkyl group as defined hereinwherein all of the hydrogen atoms are independently replaced by ahalogen, e.g., fluoro, bromo, chloro, or iodo. In some embodiments, thealkyl moiety has 1 to 8 carbon atoms (“C₁₋₈ perhaloalkyl”). In someembodiments, the alkyl moiety has 1 to 6 carbon atoms (“C₁₋₆perhaloalkyl”). In some embodiments, the alkyl moiety has 1 to 4 carbonatoms (“C₁₋₄ perhaloalkyl”). In some embodiments, the alkyl moiety has 1to 3 carbon atoms (“C₁₋₃ perhaloalkyl”). In some embodiments, the alkylmoiety has 1 to 2 carbon atoms (“C₁₋₂ perhaloalkyl”). In someembodiments, all of the hydrogen atoms are replaced with fluoro. In someembodiments, all of the hydrogen atoms are replaced with chloro.Examples of perhaloalkyl groups include —CF₃, —CF₂CF₃, —CF₂CF₂CF₃,—CCl₃, —CFCl₂, —CF₂Cl, and the like.

The term “alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, and one or morecarbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds). In someembodiments, an alkenyl group has 2 to 20 carbon atoms (“C₂₋₂₀alkenyl”). In some embodiments, an alkenyl group has 2 to 12 carbonatoms (“C₂₋₁₂ alkenyl”). In some embodiments, an alkenyl group has 2 to11 carbon atoms (“C₂₋₁₁ alkenyl”). In some embodiments, an alkenyl grouphas 2 to 10 carbon atoms (“C₂₋₁₀ alkenyl”). In some embodiments, analkenyl group has 2 to 9 carbon atoms (“C₂₋₉ alkenyl”). In someembodiments, an alkenyl group has 2 to 8 carbon atoms (“C₂₋₈ alkenyl”).In some embodiments, an alkenyl group has 2 to 7 carbon atoms (“C₂₋₇alkenyl”). In some embodiments, an alkenyl group has 2 to 6 carbon atoms(“C₂₋₆ alkenyl”). In some embodiments, an alkenyl group has 2 to 5carbon atoms (“C₂₋₅ alkenyl”). In some embodiments, an alkenyl group has2 to 4 carbon atoms (“C₂₋₄ alkenyl”). In some embodiments, an alkenylgroup has 2 to 5 carbon atoms (“C₂₋₃ alkenyl”). In some embodiments, analkenyl group has 2 carbon atoms (“C₂ alkenyl”). The one or morecarbon-carbon double bonds can be internal (such as in 2-butenyl) orterminal (such as in 1-butenyl). Examples of C₂₋₄ alkenyl groups includeethenyl (C₂), 1-propenyl (C₃), 2-propenyl (C₃), 1-butenyl (C₄),2-butenyl (C₄), butadienyl (C₄), and the like. Examples of C₂₋₆ alkenylgroups include the aforementioned C₂₋₄ alkenyl groups as well aspentenyl (C₅), pentadienyl (C₅), hexenyl (C), and the like. Additionalexamples of alkenyl include heptenyl (C₇), octenyl (C₈), octatrienyl(C₈), and the like. Unless otherwise specified, each instance of analkenyl group is independently optionally unsubstituted (an“unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) withone or more substituents. In certain embodiments, the alkenyl group isunsubstituted C₂₋₁₀ alkenyl. In certain embodiments, the alkenyl groupis substituted C₂₋₁₀ alkenyl. In an alkenyl group, a C═C double bond forwhich the stereochemistry is not specified (e.g., —CH═CHCH₃ or

may be in the (E)- or (Z)-configuration.

The term “alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms, and one or morecarbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds). In someembodiments, an alkynyl group has 2 to 10 carbon atoms (“C₂₋₁₀alkynyl”). In some embodiments, an alkynyl group has 2 to 9 carbon atoms(“C₂₋₉ alkynyl”). In some embodiments, an alkynyl group has 2 to 8carbon atoms (“C₂₋₈ alkynyl”). In some embodiments, an alkynyl group has2 to 7 carbon atoms (“C₂₋₇ alkynyl”). In some embodiments, an alkynylgroup has 2 to 6 carbon atoms (“C₂₋₆ alkynyl”). In some embodiments, analkynyl group has 2 to 5 carbon atoms (“C₂₋₅ alkynyl”). In someembodiments, an alkynyl group has 2 to 4 carbon atoms (“C₂₋₄ alkynyl”).In some embodiments, an alkynyl group has 2 to 3 carbon atoms (“C₂₋₃alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“C₂alkynyl”). The one or more carbon-carbon triple bonds can be internal(such as in 2-butynyl) or terminal (such as in 1-butynyl). Examples ofC₂₋₄ alkynyl groups include ethynyl (C₂), 1-propynyl (C₃), 2-propynyl(C₃), 1-butynyl (C₄), 2-butynyl (C₄), and the like. Examples of C₂₋₆alkenyl groups include the aforementioned C₂₋₄ alkynyl groups as well aspentynyl (C₅), hexynyl (C₆), and the like. Additional examples ofalkynyl include heptynyl (C₇), octynyl (C₈), and the like. Unlessotherwise specified, each instance of an alkynyl group is independentlyoptionally substituted, e.g., unsubstituted (an “unsubstituted alkynyl”)or substituted (a “substituted alkynyl”) with one or more substituents.In certain embodiments, the alkynyl group is unsubstituted C₂₋₁₀alkynyl. In certain embodiments, the alkynyl group is substituted C₂₋₁₀alkynyl.

The term “Carbocyclyl” or “carbocyclic” refers to a radical of anon-aromatic cyclic hydrocarbon group having from 3 to 13 ring carbonatoms (“C₃₋₁₃ carbocyclyl”) and zero heteroatoms in the non-aromaticring system. In some embodiments, a carbocyclyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ carbocyclyl”). In some embodiments, a carbocyclylgroup has 3 to 7 ring carbon atoms (“C₃₋₇ carbocyclyl”). In someembodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C₃₋₆carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclyl groupsinclude cyclopropyl (C₃), cyclopropenyl (C₃), cyclobutyl (C₄),cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅), cyclohexyl(C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and the like. ExemplaryC₃₋₈ carbocyclyl groups include the aforementioned C₃₋₆ carbocyclylgroups as well as cycloheptyl (C₇), cycloheptenyl (C₇), cycloheptadienyl(C₇), cycloheptatrienyl (C₇), cyclooctyl (C₈), cyclooctenyl (C₈),bicyclo[2.2.1]heptanyl (C₇), bicyclo[2.2.2]octanyl (C₈), and the like.Exemplary C₃₋₁₀ carbocyclyl groups include the aforementioned C₃₋₈carbocyclyl groups as well as cyclononyl (C₉), cyclononenyl (C₉),cyclodecyl (C₁₀), cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉),decahydronaphthalenyl (C₁₀), spiro[4.5]decanyl (C₁₀), and the like. Asthe foregoing examples illustrate, in certain embodiments, thecarbocyclyl group is either monocyclic (“monocyclic carbocyclyl”) orcontain a fused, bridged, or spiro ring system such as a bicyclic system(“bicyclic carbocyclyl”). Carbocyclyl can be saturated, and saturatedcarbocyclyl is referred to as “cycloalkyl.” In some embodiments,carbocyclyl is a monocyclic, saturated carbocyclyl group having from 3to 10 ring carbon atoms (“C₃₋₁₀ cycloalkyl”). In some embodiments, acycloalkyl group has 3 to 8 ring carbon atoms (“C₃₋₈ cycloalkyl”). Insome embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms (“C₃₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ringcarbon atoms (“C₅₋₆ cycloalkyl”). In some embodiments, a cycloalkylgroup has 5 to 10 ring carbon atoms (“C₅₋₁₀ cycloalkyl”). Examples ofC₅₋₆ cycloalkyl groups include cyclopentyl (C₅) and cyclohexyl (C₅).Examples of C₃₋₆ cycloalkyl groups include the aforementioned C₅₋₆cycloalkyl groups as well as cyclopropyl (C₃) and cyclobutyl (C₄).Examples of C₃₋₈ cycloalkyl groups include the aforementioned C₃₋₆cycloalkyl groups as well as cycloheptyl (C₇) and cyclooctyl (C₈).Unless otherwise specified, each instance of a cycloalkyl group isindependently unsubstituted (an “unsubstituted cycloalkyl”) orsubstituted (a “substituted cycloalkyl”) with one or more substituents.In certain embodiments, the cycloalkyl group is unsubstituted C₃₋₁₀cycloalkyl. In certain embodiments, the cycloalkyl group is substitutedC₃₋₁₀ cycloalkyl. Carbocyclyl can be partially unsaturated. Carbocyclylmay include zero, one, or more (e.g., two, three, or four, as valencypermits) C═C double bonds in all the rings of the carbocyclic ringsystem that are not aromatic or heteroaromatic. Carbocyclyl includingone or more (e.g., two or three, as valency permits) C═C double bonds inthe carbocyclic ring is referred to as “cycloalkenyl.” Carbocyclylincluding one or more (e.g., two or three, as valency permits) C≡ triplebonds in the carbocyclic ring is referred to as “cycloalkynyl.”Carbocyclyl includes aryl. “Carbocyclyl” also includes ring systemswherein the carbocyclyl ring, as defined above, is fused with one ormore aryl or heteroaryl groups wherein the point of attachment is on thecarbocyclyl ring, and in such instances, the number of carbons continueto designate the number of carbons in the carbocyclic ring system.Unless otherwise specified, each instance of a carbocyclyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted carbocyclyl”) or substituted (a “substitutedcarbocyclyl”) with one or more substituents. In certain embodiments, thecarbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl. In certainembodiments, the carbocyclyl group is a substituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl is substituted or unsubstituted,3- to 7-membered, and monocyclic. In certain embodiments, thecarbocyclyl is substituted or unsubstituted, 5- to 13-membered, andbicyclic.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 10 ring carbon atoms (“C₃₋₁₀cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ cycloalkyl”). In someembodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ cycloalkyl”). Examples of C₅₋₆ cycloalkyl groupsinclude cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃₋₆cycloalkyl groups include the aforementioned C₅₋₆ cycloalkyl groups aswell as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃₋₈cycloalkyl groups include the aforementioned C₃₋₆ cycloalkyl groups aswell as cycloheptyl (C₇) and cyclooctyl (C₈). Unless otherwisespecified, each instance of a cycloalkyl group is independentlyunsubstituted (an “unsubstituted cycloalkyl”) or substituted (a“substituted cycloalkyl”) with one or more substituents. In certainembodiments, the cycloalkyl group is unsubstituted C₃₋₁₀ cycloalkyl. Incertain embodiments, the cycloalkyl group is substituted C₃₋₁₀cycloalkyl. In certain embodiments, the carbocyclyl includes 0, 1, or 2C═C double bonds in the carbocyclic ring system, as valency permits.

The term “Heterocyclyl” or “heterocyclic” refers to a radical of a 3- to13-membered non-aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). Inheterocyclyl groups that contain one or more nitrogen atoms, the pointof attachment can be a carbon or nitrogen atom, as valency permits. Aheterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”)or a fused, bridged, or spiro ring system such as a bicyclic system(“bicyclic heterocyclyl”). A heterocyclyl group can be saturated or canbe partially unsaturated. Heterocyclyl may include zero, one, or more(e.g., two, three, or four, as valency permits) double bonds in all therings of the heterocyclic ring system that are not aromatic orheteroaromatic. Partially unsaturated heterocyclyl groups includesheteroaryl. Heterocyclyl bicyclic ring systems can include one or moreheteroatoms in one or both rings. “Heterocyclyl” also includes ringsystems wherein the heterocyclyl ring, as defined above, is fused withone or more carbocyclyl groups wherein the point of attachment is eitheron the carbocyclyl or heterocyclyl ring, or ring systems wherein theheterocyclyl ring, as defined above, is fused with one or more aryl orheteroaryl groups, wherein the point of attachment is on theheterocyclyl ring, and in such instances, the number of ring memberscontinue to designate the number of ring members in the heterocyclylring system. Unless otherwise specified, each instance of heterocyclylis independently optionally substituted, e.g., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substitutedheterocyclyl”) with one or more substituents. In certain embodiments,the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. Incertain embodiments, the heterocyclyl group is substituted 3-10 memberedheterocyclyl. In certain embodiments, the heterocyclyl is substituted orunsubstituted, 3- to 7-membered, and monocyclic. In certain embodiments,the heterocyclyl is substituted or unsubstituted, 5- to 13-membered, andbicyclic.

In some embodiments, a heterocyclyl group is a 5-10 memberednon-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-10 membered heterocyclyl”). In someembodiments, a heterocyclyl group is a 5-8 membered non-aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms, wherein eachheteroatom is independently selected from nitrogen, oxygen, and sulfur(“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl groupis a 5-6 membered non-aromatic ring system having ring carbon atoms and1-4 ring heteroatoms, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In someembodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatomsselected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6membered heterocyclyl has 1-2 ring heteroatoms selected from nitrogen,oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclylhas one ring heteroatom selected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude aziridinyl, oxiranyl, or thiiranyl. Exemplary 4-memberedheterocyclyl groups containing one heteroatom include azetidinyl,oxetanyl and thietanyl. Exemplary 5-membered heterocyclyl groupscontaining one heteroatom include tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyland pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include dioxolanyl, oxasulfuranyl,disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclylgroups containing three heteroatoms include triazolinyl, oxadiazolinyl,and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containingone heteroatom include piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include piperazinyl, morpholinyl, dithianyl, and dioxanyl.Exemplary 6-membered heterocyclyl groups containing two heteroatomsinclude triazinyl. Exemplary 7-membered heterocyclyl groups containingone heteroatom include azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom includeazocanyl, oxecanyl, and thiocanyl. Exemplary 5-membered heterocyclylgroups fused to a C₆ aryl ring (also referred to herein as a5,6-bicyclic heterocyclic ring) include indolinyl, isoindolinyl,dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl, and thelike. Exemplary 6-membered heterocyclyl groups fused to an aryl ring(also referred to herein as a 6,6-bicyclic heterocyclic ring) include,without limitation, decahydroquinolinyl, decahydroisoquinolinyl,octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl,decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl,phthalimidyl, naphthalimidyl, chromanyl, chromenyl,1H-benzo[e][1,4]diazepinyl, 1,4,5,7-tetrahydropyrano[3,4-b]pyrrolyl,5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl,5,7-dihydro-4H-thieno[2,3-c]pyranyl,2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl,4,5,6,7-tetrahydro-1H-pyrrolo[2,3-b]pyridinyl,4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl,4,5,6,7-tetrahydrothieno[3,2-b]pyridinyl,1,2,3,4-tetrahydro-1,6-naphthyridinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and the like.

The term “Aryl” refers to a radical of a monocyclic or polycyclic (e.g.,bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or14 π-electrons shared in a cyclic array) having 6-14 ring carbon atomsand zero heteroatoms provided in the aromatic ring system (“C₆₋₁₄aryl”). In some embodiments, an aryl group has six ring carbon atoms(“C₆ aryl”; e.g., phenyl). In some embodiments, an aryl group has tenring carbon atoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and2-naphthyl). In some embodiments, an aryl group has fourteen ring carbonatoms (“C₁₄ aryl”; e.g., anthracyl). “Aryl” also includes ring systemswherein the aryl ring, as defined above, is fused with one or morecarbocyclyl or heterocyclyl groups wherein the radical or point ofattachment is on the aryl ring, and in such instances, the number ofcarbon atoms continue to designate the number of carbon atoms in thearyl ring system. Unless otherwise specified, each instance of an arylgroup is independently optionally substituted, e.g., unsubstituted (an“unsubstituted aryl”) or substituted (a “substituted aryl”) with one ormore substituents. In certain embodiments, the aryl group isunsubstituted C₆₋₁₄ aryl. In certain embodiments, the aryl group issubstituted C₆₋₁₄ aryl.

The term “aralkyl” is a subset of “alkyl” and refers to an alkyl groupsubstituted by an aryl group, wherein the point of attachment is on thealkyl moiety.

The term “heteroaryl” refers to a radical of a 5-10 membered monocyclicor bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 □ electronsshared in a cyclic array) having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, e.g., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl). In certain embodiments, theheteroaryl is substituted or unsubstituted, 5- or 6-membered, monocyclicheteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ring systemare independently oxygen, nitrogen, or sulfur. In certain embodiments,the heteroaryl is substituted or unsubstituted, 9- or 10-membered,bicyclic heteroaryl, wherein 1, 2, 3, or 4 atoms in the heteroaryl ringsystem are independently oxygen, nitrogen, or sulfur.

In some embodiments, a heteroaryl group is a 5-10 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsselected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6membered heteroaryl has 1-2 ring heteroatoms selected from nitrogen,oxygen, and sulfur. In some embodiments, the 5-6 membered heteroaryl has1 ring heteroatom selected from nitrogen, oxygen, and sulfur. Unlessotherwise specified, each instance of a heteroaryl group isindependently optionally substituted, e.g., unsubstituted(“unsubstituted heteroaryl”) or substituted (“substituted heteroaryl”)with one or more substituents. In certain embodiments, the heteroarylgroup is unsubstituted 5-14 membered heteroaryl. In certain embodiments,the heteroaryl group is substituted 5-14 membered heteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatom includepyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groupscontaining two heteroatoms include imidazolyl, pyrazolyl, oxazolyl,isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroarylgroups containing three heteroatoms include triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include tetrazolyl. Exemplary 6-membered heteroaryl groupscontaining one heteroatom include pyridinyl. Exemplary 6-memberedheteroaryl groups containing two heteroatoms include pyridazinyl,pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groupscontaining three or four heteroatoms include triazinyl and tetrazinyl,respectively. Exemplary 7-membered heteroaryl groups containing oneheteroatom include azepinyl, oxepinyl, and thiepinyl. Exemplary5,6-bicyclic heteroaryl groups include indolyl, isoindolyl, indazolyl,benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl,benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl,benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl,indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl,cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplarytricyclic heteroaryl groups include phenanthridinyl, dibenzofuranyl,carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, and phenazinyl.

The term “heteroaralkyl” is a subset of “alkyl” and refers to an alkylgroup substituted by a heteroaryl group, wherein the attachment is onthe alkyl moiety.

The term “unsaturated” or “partially unsaturated” refers to a moietythat includes at least one double or triple bond. The term “partiallyunsaturated” is intended to encompass rings having multiple sites ofunsaturation, but is not intended to include aromatic groups (e.g., arylor heteroaryl groups) as herein defined. Likewise, “saturated” refers toa group that does not contain a double or triple bond, i.e., containsall single bonds.

In some embodiments, aliphatic, alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aryl, and heteroaryl groups, as defined herein, areoptionally substituted (e.g., “substituted” or “unsubstituted” alkyl,“substituted” or “unsubstituted” alkenyl, “substituted” or“unsubstituted” alkynyl, “substituted” or “unsubstituted” carbocyclyl,“substituted” or “unsubstituted” heterocyclyl, “substituted” or“unsubstituted” aryl or “substituted” or “unsubstituted” heteroarylgroup). In general, the term “substituted”, whether preceded by the term“optionally” or not, means that at least one hydrogen present on a group(e.g., a carbon or nitrogen atom) is replaced with a permissiblesubstituent, e.g., a substituent which upon substitution results in astable compound, e.g., a compound which does not spontaneously undergotransformation such as by rearrangement, cyclization, elimination, orother reaction. Unless otherwise indicated, a “substituted” group has asubstituent at one or more substitutable positions of the group, andwhen more than one position in any given structure is substituted, thesubstituent is either the same or different at each position. The term“substituted” is contemplated to include substitution with allpermissible substituents of organic compounds, any of the substituentsdescribed herein that results in the formation of a stable compound. Thepresent disclosure contemplates any and all such combinations in orderto arrive at a stable compound. For purposes of this disclosure,heteroatoms such as nitrogen may have hydrogen substituents and/or anysuitable substituent as described herein which satisfy the valencies ofthe heteroatoms and results in the formation of a stable moiety.

A group is optionally substituted unless expressly provided otherwise.The term “optionally substituted” refers to being substituted orunsubstituted.

In certain embodiments, alkyl, alkenyl, alkynyl, heteroalkyl,heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl groups are optionally substituted (e.g., “substituted” or“unsubstituted” alkyl, “substituted” or “unsubstituted” alkenyl,“substituted” or “unsubstituted” alkynyl, heteroalkyl, “substituted” or“unsubstituted” heteroalkenyl, “substituted” or “unsubstituted”heteroalkynyl, “substituted” or “unsubstituted”, “substituted” or“unsubstituted” carbocyclyl, “substituted” or “unsubstituted”heterocyclyl, “substituted” or “unsubstituted” aryl or “substituted” or“unsubstituted” heteroaryl group). In general, the term “substituted”,means that at least one hydrogen present on a group is replaced with apermissible substituent, e.g., a substituent which upon substitutionresults in a stable compound, e.g., a compound which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, or other reaction. Unless otherwise indicated,a “substituted” group has a substituent at one or more substitutablepositions of the group, and when more than one position in any givenstructure is substituted, the substituent is either the same ordifferent at each position. The term “substituted” is contemplated toinclude substitution with all permissible substituents of organiccompounds, and includes any of the substituents described herein thatresults in the formation of a stable compound. The present disclosurecontemplates any and all such combinations in order to arrive at astable compound. For purposes of this disclosure, heteroatoms such asnitrogen may have hydrogen substituents and/or any suitable substituentas described herein which satisfy the valencies of the heteroatoms andresults in the formation of a stable moiety.

Exemplary carbon atom substituents include halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂, —N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻,—N(OR^(cc))R^(bb), —SH, —SR—, —SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO,—C(OR^(cc))₂, —CO₂R^(aa), —OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂,—OC(═O)N(R^(bb))₂, —NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa),—NR^(bb)C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa),—OC(═NR^(bb))R^(aa), —OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—OC(═NR^(bb))N(R^(bb))₂, —NR^(bb)C(═NR^(bb))N(R^(bb))₂,—C(═O)NR^(bb)SO₂R^(aa), —NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa),—SO₂OR^(aa), —OSO₂R^(aa), —S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃,—OSi(R^(aa))₃—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa),—SC(═S)SR^(aa), —SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa),—SC(═O)R^(aa), —P(═O)(R^(cc))₂, —P(═O)(OR^(cc))₂, —OP(═O)(R^(aa))₂,—OP(═O)(OR^(cc))₂, —P(═O)(N(R^(bb))₂)₂, —OP(═O)(N(R^(bb))₂)₂,—NR^(bb)P(═O)(R^(aa))₂, —NR^(bb)P(═O)(OR^(cc))₂,—NR^(bb)P(═O)(N(R^(bb))₂)₂, —P(R^(cc))₂, —P(OR^(cc))₂, —P(R^(cc))₃ ⁺X⁻,—P(OR^(cc))₃ ⁺X⁻, —P(R^(cc))₄, —P(OR^(cc))₄, —OP(R^(cc))₂, —OP(R^(cc))₃⁺X⁻, —OP(OR^(cc))₂, —OP(OR^(cc))₃ ⁺X⁻, —OP(R^(cc))₄, —OP(OR^(cc))₄,—B(R^(aa))₂, —B(OR^(cc))₂, —BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl, C₁₋₁₀perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, heteroC₁₋₁₀ alkyl,heteroC₂₋₁₀ alkenyl, heteroC₂₋₁₀ alkynyl, C₁₋₁₀ carbocyclyl, 3-14membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups; wherein X⁻ is acounterion;

-   -   or two geminal hydrogens on a carbon atom are replaced with the        group ═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R—,        ═NNR^(bb)C(═O)OR^(aa), ═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or        ═NOR^(cc); each instance of R^(aa) is, independently, selected        from C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀        alkynyl, heteroC₁₋₁₀ alkyl, heteroC₂₋₁₀ alkenyl, heteroC₂₋₁₀        alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄        aryl, and 5-14 membered heteroaryl, or two R^(aa) groups are        joined to form a 3-14 membered heterocyclyl or 5-14 membered        heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,        heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl,        heterocyclyl, aryl, and heteroaryl is independently substituted        with 0, 1, 2, 3, 4, or 5 R^(dd) groups;    -   each instance of R^(bb) is, independently, selected from        hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),        —C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(cc))OR^(aa),        —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(aa), —SO₂OR^(cc),        —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),        —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, —P(═O)(N(R^(cc))₂)₂, C₁₋₁₀        alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl,        heteroC₁₋₁₀ alkyl, heteroC₂₋₁₀ alkenyl, heteroC₂₋₁₀ alkynyl,        C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and        5-14 membered heteroaryl, or two R^(bb) groups are joined to        form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl        ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl,        heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl,        and heteroaryl is independently substituted with 0, 1, 2, 3, 4,        or 5 R^(dd) groups; wherein X⁻ is a counterion;    -   each instance of R^(cc) is, independently, selected from        hydrogen, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀        alkynyl, heteroC₁₋₁₀ alkyl, heteroC₂₋₁₀ alkenyl, heteroC₂₋₁₀        alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄        aryl, and 5-14 membered heteroaryl, or two R^(cc) groups are        joined to form a 3-14 membered heterocyclyl or 5-14 membered        heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,        heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl,        heterocyclyl, aryl, and heteroaryl is independently substituted        with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(dd) is, independently, selected from halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —ON(R^(ff))₂, —N(R^(ff))₂,—N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff), —SH, —SR^(ee), —SSR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NR^(ff)C(═O)N(R^(ff))₂, —C(═NR^(ff))OR^(ee),—OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee), —C(═NR^(ff))N(R^(ff))₂,—OC(═NR^(ff))N(R^(ff))₂, —NR^(ff)C(═NR^(ff))N(R^(ff))₂,—NR^(ff)SO₂R^(ee), —SO₂N(R^(ff))₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),—S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,—C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)(OR^(ee))₂,—P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, heteroC₁₋₆alkyl,heteroC₂₋₆alkenyl, heteroC₂₋₆alkynyl, C₃₋₁₀ carbocyclyl, 3-10 memberedheterocyclyl, C₆₋₁₀ aryl, 5-10 membered heteroaryl, wherein each alkyl,alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups, or two geminalR^(dd) substituents can be joined to form ═O or ═S; wherein X⁻ is acounterion;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, heteroC₁₋₆ alkyl,heteroC₂₋₆alkenyl, heteroC₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀ aryl,3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein eachalkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups;

each instance of R^(ff) is, independently, selected from hydrogen, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, heteroC₁₋₆alkyl,heteroC₂₋₆alkenyl, heteroC₂₋₆alkynyl, C₃₋₁₀ carbocyclyl, 3-10 memberedheterocyclyl, C₆₋₁₀ aryl and 5-10 membered heteroaryl, or two R^(ff)groups are joined to form a 3-10 membered heterocyclyl or 5-10 memberedheteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl,heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups; and each instance of R^(gg) is, independently, halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆alkyl)₂, —N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)+X⁻,—NH₃ ⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁-6 alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃ —C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)(OC₁₋₆alkyl)₂, —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁_6 perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,heteroC₁₋₆alkyl, heteroC₂-6 alkenyl, heteroC₂₋₆alkynyl, C₃₋₁₀carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R⁹⁹ substituents can be joined to form ═O or═S; wherein X⁻ is a counterion.

In certain embodiments, the carbon atom substituents are independentlyhalogen, substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, —OR^(aa), —SR^(aa), —N(R^(bb))₂, —CN, —SCN,—NO₂, —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)R^(aa),—OCO₂R^(aa), —OC(═O)N(R^(bb))₂, —NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa),or —NR^(bb)C(═O)N(R^(bb))₂. In certain embodiments, the carbon atomsubstituents are independently halogen, substituted (e.g., substitutedwith one or more halogen) or unsubstituted C₁₋₆ alkyl, —OR^(aa),SR^(aa), —N(R^(bb))₂, —CN, —SCN, —NO₂, —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, —OC(═O)R^(aa), —OCO₂R^(aa), —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), or —NR^(bb)C(═O)N(R^(bb))₂,wherein R^(aa) is hydrogen, substituted (e.g., substituted with one ormore halogen) or unsubstituted C₁₋₆ alkyl, an oxygen protecting groupwhen attached to an oxygen atom, or a sulfur protecting group (e.g.,acetamidomethyl, t-Bu, 3-nitro-2-pyridine sulfenyl, 2-pyridine-sulfenyl,or triphenylmethyl) when attached to a sulfur atom; and each R^(bb) isindependently hydrogen, substituted (e.g., substituted with one or morehalogen) or unsubstituted C₁₋₆ alkyl, or a nitrogen protecting group. Incertain embodiments, the carbon atom substituents are independentlyhalogen, substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, —OR^(aa), —SR^(aa), —N(R^(bb))₂, —CN, —SCN, or—NO₂. In certain embodiments, the carbon atom substituents areindependently halogen, substituted (e.g., substituted with one or morehalogen moieties) or unsubstituted C₁₋₆ alkyl, —OR^(aa), —SR^(aa),—N(R^(bb))₂, —CN, —SCN, or —NO₂, wherein R^(aa) is hydrogen, substituted(e.g., substituted with one or more halogen) or unsubstituted C₁₋₆alkyl, an oxygen protecting group when attached to an oxygen atom, or asulfur protecting group (e.g., acetamidomethyl, t-Bu, 3-nitro-2-pyridinesulfenyl, 2-pyridine-sulfenyl, or triphenylmethyl) when attached to asulfur atom; and each R^(bb) is independently hydrogen, substituted(e.g., substituted with one or more halogen) or unsubstituted C₁₋₆alkyl, or a nitrogen protecting group.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a positively charged group in order to maintainelectronic neutrality. An anionic counterion may be monovalent (i.e.,including one formal negative charge). An anionic counterion may also bemultivalent (i.e., including more than one formal negative charge), suchas divalent or trivalent. Exemplary counterions include halide ions(e.g., F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HCO₃ ⁻, HSO₄ ⁻,sulfonate ions (e.g., methansulfonate, trifluoromethanesulfonate,p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate,naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate,ethan-1-sulfonic acid-2-sulfonate, and the like), carboxylate ions(e.g., acetate, propanoate, benzoate, glycerate, lactate, tartrate,glycolate, gluconate, and the like), BF₄ ⁻, PF₄ ⁻, PF₆ ⁻, AsF₆ ⁻, SbF₆⁻, B[3,5-(CF₃)₂C₆H₃]₄]⁻, B(C₆F₅)₄ ⁻, BPh₄ ⁻, Al(OC(CF₃)₃)₄, andcarborane anions (e.g., CB₁₁H₁₂ ⁻ or (HCB₁₁Me₅Br₆)⁻). Exemplarycounterions which may be multivalent include CO₃ ²⁻, HPO₄ ²⁻, PO₄ ³⁻,B₄O₇ ²⁻, SO₄ ⁴⁻, S₂O₃ ²⁻, carboxylate anions (e.g., tartrate, citrate,fumarate, maleate, malate, malonate, gluconate, succinate, glutarate,adipate, pimelate, suberate, azelate, sebacate, salicylate, phthalates,aspartate, glutamate, and the like), and carboranes.

“Halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro,—Cl), bromine (bromo, —Br), or iodine (iodo, —I).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quaternary nitrogen atoms.Exemplary nitrogen atom substituents include hydrogen, —OH, —OR^(aa),—N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(bb))R—, —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂,—SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂,—C(═O)SR^(cc), —C(═S)SR^(cc), —P(═O)(OR^(cc))₂, —P(═O)(R^(cc))₂,—P(═O)(N(R^(cc))₂)₂, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl,C₂₋₁₀ alkynyl, heteroC₁₋₁₀ alkyl, heteroC₂₋₁₀ alkenyl, heteroC₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(cc) groups attached to an N atom arejoined to form a 3-14 membered heterocyclyl or 5-14 membered heteroarylring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl,heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups, andwherein R^(aa), R^(bb), R^(cc) and R^(dd) are as defined above.

In certain embodiments, each nitrogen atom substituent is independentlysubstituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂, ora nitrogen protecting group. In certain embodiments, each nitrogen atomsubstituent is independently substituted (e.g., substituted with one ormore halogen) or unsubstituted C₁₋₁₀ alkyl, —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, or a nitrogen protecting group, wherein R^(aa) ishydrogen, substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₁₀ alkyl, or an oxygen protecting group when attachedto an oxygen atom; and each R^(bb) is independently hydrogen,substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₁₀ alkyl, or a nitrogen protecting group. In certainembodiments, each nitrogen atom substituent is independently substituted(e.g., substituted with one or more halogen) or unsubstituted C₁₋₆ alkylor a nitrogen protecting group.

In certain embodiments, the nitrogen atom substituents are independentlysubstituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂, ora nitrogen protecting group. In certain embodiments, the nitrogen atomsubstituents are independently substituted (e.g., substituted with oneor more halogen) or unsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, or a nitrogen protecting group, wherein R^(aa) ishydrogen, substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, or an oxygen protecting group when attached toan oxygen atom; and each R^(bb) is independently hydrogen, substituted(e.g., substituted with one or more halogen) or unsubstituted C₁₋₆alkyl, or a nitrogen protecting group. In certain embodiments, thenitrogen atom substituents are independently substituted (e.g.,substituted with one or more halogen) or unsubstituted C₁₋₆ alkyl or anitrogen protecting group.

In certain embodiments, the substituent present on a nitrogen atom is anitrogen protecting group (also referred to as an amino protectinggroup). Nitrogen protecting groups include —OH, —OR^(aa), —N(R^(cc))₂,—C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa),—C(═NR^(cc))R^(aa), —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂,—SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂,—C(═O)SR^(cc), —C(═S)SR^(cc), C₁₋₁₀ alkyl (e.g., aralkyl,heteroaralkyl), C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl, 3-14membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl groups,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,aralkyl, aryl, and heteroaryl is independently substituted with 0, 1, 2,3, 4, or 5 R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc), and R^(dd)are as defined herein. Nitrogen protecting groups are well known in theart and include those described in detail in Protecting Groups inOrganic Synthesis, T. W. Greene and P. G. M. Wuts, 3^(rd) edition, JohnWiley & Sons, 1999, incorporated herein by reference.

Amide nitrogen protecting groups (e.g., —C(═O)R^(aa)) include formamide,acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide,phenylacetamide, 3-phenylpropanamide, picolinamide,3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide,p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide,acetoacetamide, (N′-dithiobenzyloxyacylamino)acetamide,3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide,2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine,o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Carbamate nitrogen protecting groups (e.g., —C(═O)OR^(aa)) includemethyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc),9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethylcarbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzylcarbamate.

Sulfonamide nitrogen protecting groups (e.g., —S(═O)₂R^(aa)) includep-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), (3-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include phenothiazinyl-(10)-acylderivative, N′-p-toluenesulfonylaminoacyl derivative,N′-phenylaminothioacyl derivative, N-benzoylphenylalanyl derivative,N-acetylmethionine derivative, 4,5-diphenyl-3-oxazolin-2-one,N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide,N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentaneadduct (STABASE), 5-substituted1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate,N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys). Insome embodiments, two instances of a nitrogen protecting group togetherwith the nitrogen atoms to which the nitrogen protecting groups areattached are N,N′-isopropylidenediamine. In certain embodiments, anitrogen protecting group is Bn, Boc, Cbz, Fmoc, trifluoroacetyl,triphenylmethyl, acetyl, or Ts.

In certain embodiments, the oxygen atom substituents are independentlysubstituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂, oran oxygen protecting group. In certain embodiments, the oxygen atomsubstituents are independently substituted (e.g., substituted with oneor more halogen) or unsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, or an oxygen protecting group, wherein R^(aa) ishydrogen, substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, or an oxygen protecting group when attached toan oxygen atom; and each R^(bb) is independently hydrogen, substituted(e.g., substituted with one or more halogen) or unsubstituted C₁₋₆alkyl, or a nitrogen protecting group. In certain embodiments, theoxygen atom substituents are independently substituted (e.g.,substituted with one or more halogen) or unsubstituted C₁₋₆ alkyl or anoxygen protecting group.

In certain embodiments, the substituent present on an oxygen atom is anoxygen protecting group (also referred to herein as an “hydroxylprotecting group”). Oxygen protecting groups include —R^(aa),—N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃ ⁺X⁻,—P(OR^(cc))₂, —P(OR^(cc))₃ ⁺X⁻, —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, and—P(═O)(N(R^(bb))₂)₂, wherein X⁻, R^(aa), R^(bb), and R^(cc) are asdefined herein. Oxygen protecting groups are well known in the art andinclude those described in detail in Protecting Groups in OrganicSynthesis, T. W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley &Sons, 1999, incorporated herein by reference.

Exemplary oxygen protecting groups include methyl, methoxymethyl (MOM),methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate,9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate(TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec),2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutylcarbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′,N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate,borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate,sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate(Ts).

In certain embodiments, an oxygen protecting group is silyl, TBDPS,TBDMS, TIPS, TES, TMS, MOM, THP, t-Bu, Bn, allyl, acetyl, pivaloyl, orbenzoyl.

In certain embodiments, the sulfur atom substituents are independentlysubstituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂, ora sulfur protecting group. In certain embodiments, the sulfur atomsubstituents are independently substituted (e.g., substituted with oneor more halogen) or unsubstituted C₁₋₆ alkyl, —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, or a sulfur protecting group, wherein R^(aa) ishydrogen, substituted (e.g., substituted with one or more halogen) orunsubstituted C₁₋₆ alkyl, or an oxygen protecting group when attached toan oxygen atom; and each R^(bb) is independently hydrogen, substituted(e.g., substituted with one or more halogen) or unsubstituted C₁₋₆alkyl, or a nitrogen protecting group. In certain embodiments, thesulfur atom substituents are independently substituted (e.g.,substituted with one or more halogen) or unsubstituted C₁₋₆ alkyl or asulfur protecting group.

In certain embodiments, the substituent present on a sulfur atom is asulfur protecting group (also referred to as a “thiol protectinggroup”). In some embodiments, each sulfur protecting group is selectedfrom the group consisting of —R^(aa), —N(R^(bb))₂, —C(═O)SR^(aa),—C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa),—C(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —S(═O)R^(aa), —SO₂R^(aa),—Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃ ⁺X⁻, —P(OR^(cc))₂, —P(OR^(cc))₃⁺X⁻, —P(═O)(R^(cc))₂, —P(═O)(OR^(cc))₂, and —P(═O)(N(R^(bb))₂)₂, whereinR^(aa), R^(bb), and R^(cc) are as defined herein. Sulfur protectinggroups are well known in the art and include those described in detailin Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.Wuts, 3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference. In certain embodiments, a sulfur protecting group isacetamidomethyl, t-Bu, 3-nitro-2-pyridine sulfenyl, 2-pyridine-sulfenyl,or triphenylmethyl.

The “molecular weight” of —R, wherein —R is any monovalent moiety, iscalculated by subtracting the atomic weight of a hydrogen atom from themolecular weight of the molecule R—H. The “molecular weight” of -L-,wherein -L- is any divalent moiety, is calculated by subtracting thecombined atomic weight of two hydrogen atoms from the molecular weightof the molecule H-L-H.

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, Figures, and Claims. The presentdisclosure is not intended to be limited in any manner by the aboveexemplary listing of substituents.

“Pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and other animals without undue toxicity,irritation, allergic response, and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, Berge et al., describepharmaceutically acceptable salts in detail in J. PharmaceuticalSciences (1977) 66:1-19. Pharmaceutically acceptable salts of thecompounds describe herein include those derived from suitable inorganicand organic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid, or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representativealkali or alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, quaternary salts.

The term “solvate” refers to forms of the compound, or a salt thereof,that are associated with a solvent, usually by a solvolysis reaction.This physical association may include hydrogen bonding. Conventionalsolvents include water, methanol, ethanol, acetic acid, DMSO, THF,diethyl ether, and the like. The compounds of Formula (I) may beprepared, e.g., in crystalline form, and may be solvated. Suitablesolvates include pharmaceutically acceptable solvates and furtherinclude both stoichiometric solvates and non-stoichiometric solvates. Incertain instances, the solvate will be capable of isolation, forexample, when one or more solvent molecules are incorporated in thecrystal lattice of a crystalline solid. “Solvate” encompasses bothsolution-phase and isolable solvates. Representative solvates includehydrates, ethanolates, and methanolates.

The term “stoichiometric solvate” refers to a solvate, which comprises acompound (e.g., a compound disclosed herein) and a solvent, wherein thesolvent molecules are an integral part of the crystal lattice, in whichthey interact strongly with the compound and each other. The removal ofthe solvent molecules will cause instability of the crystal network,which subsequently collapses into an amorphous phase or recrystallizesas a new crystalline form with reduced solvent content.

The term “non-stoichiometric solvate” refers to a solvate, whichcomprises a compound (e.g., a compound disclosed herein) and a solvent,wherein the solvent content may vary without major changes in thecrystal structure. The amount of solvent in the crystal lattice onlydepends on the partial pressure of solvent in the surroundingatmosphere. In the fully solvated state, non-stoichiometric solvatesmay, but not necessarily have to, show an integer molar ratio of solventto the compound. During drying of a non-stoichiometric solvate, aportion of the solvent may be removed without significantly disturbingthe crystal network, and the resulting solvate can subsequently beresolvated to give the initial crystalline form. Unlike stoichiometricsolvates, the desolvation and resolvation of non-stoichiometric solvatesis not accompanied by a phase transition, and all solvation statesrepresent the same crystal form.

The term “hydrate” refers to a compound that is associated with water.Typically, the number of the water molecules contained in a hydrate of acompound is in a definite ratio to the number of the compound moleculesin the hydrate. Therefore, a hydrate of a compound may be represented,for example, by the general formula R.x H₂O, wherein R is the compoundand wherein x is a number greater than 0. A given compound may form morethan one type of hydrates, including, e.g., monohydrates (x is 1), lowerhydrates (x is a number greater than 0 and smaller than 1, e.g.,hemihydrates (R.0.5H₂O)), and polyhydrates (x is a number greater than1, e.g., dihydrates (R.2H₂O) and hexahydrates (R.6H₂O)).

The term “tautomers” refer to compounds that are interchangeable formsof a particular compound structure, and that vary in the displacement ofhydrogen atoms and electrons. Thus, two structures may be in equilibriumthrough the movement of 7 electrons and an atom (usually H). Forexample, enols and ketones are tautomers because they are rapidlyinterconverted by treatment with either acid or base. Another example oftautomerism is the aci- and nitro-forms of phenylnitromethane, that arelikewise formed by treatment with acid or base.

Tautomeric forms may be relevant to the attainment of the optimalchemical reactivity and biological activity of a compound of interest.

The term “tautomers” or “tautomeric” refers to two or moreinterconvertible compounds resulting from at least one formal migrationof a hydrogen atom and at least one change in valency (e.g., a singlebond to a double bond, a triple bond to a single bond, or vice versa).The exact ratio of the tautomers depends on several factors, includingtemperature, solvent, and pH. Tautomerizations (i.e., the reactionproviding a tautomeric pair) may catalyzed by acid or base. Exemplarytautomerizations include keto-to-enol, amide-to-imide, lactam-to-lactim,enamine-to-imine, and enamine-to-(a different enamine) tautomerizations.

It is also to be understood that compounds that have the same molecularformula but differ in the nature or sequence of bonding of their atomsor the arrangement of their atoms in space are termed “isomers”. Isomersthat differ in the arrangement of their atoms in space are termed“stereoisomers”.

Stereoisomers that are not mirror images of one another are termed“diastereomers” and those that are non-superimposable mirror images ofeach other are termed “enantiomers”. When a compound has an asymmetriccenter, for example, it is bonded to four different groups, a pair ofenantiomers is possible. An enantiomer can be characterized by theabsolute configuration of its asymmetric center and is described by theR- and S-sequencing rules of Cahn and Prelog, or by the manner in whichthe molecule rotates the plane of polarized light and designated asdextrorotatory or levorotatory (i.e., as (+) or (−)-isomersrespectively). A chiral compound can exist as either individualenantiomer or as a mixture thereof. A mixture containing equalproportions of the enantiomers is called a “racemic mixture”.

The term “crystalline” or “crystalline form” refers to a solid formsubstantially exhibiting three-dimensional order. In certainembodiments, a crystalline form of a solid is a solid form that issubstantially not amorphous. In certain embodiments, the X-ray powderdiffraction (XRPD) pattern of a crystalline form includes one or moresharply defined peaks.

The term “amorphous” or “amorphous form” refers to a form of a solid(“solid form”), the form substantially lacking three-dimensional order.In certain embodiments, an amorphous form of a solid is a solid formthat is substantially not crystalline. In certain embodiments, the X-raypowder diffraction (XRPD) pattern of an amorphous form includes a widescattering band with a peak at 2θ of, e.g., between 20 and 70°,inclusive, using CuKα radiation. In certain embodiments, the XRPDpattern of an amorphous form further includes one or more peaksattributed to crystalline structures. In certain embodiments, themaximum intensity of any one of the one or more peaks attributed tocrystalline structures observed at a 2θ of between 20 and 70°,inclusive, is not more than 300-fold, not more than 100-fold, not morethan 30-fold, not more than 10-fold, or not more than 3-fold of themaximum intensity of the wide scattering band. In certain embodiments,the XRPD pattern of an amorphous form includes no peaks attributed tocrystalline structures.

The term “co-crystal” refers to a crystalline structure comprising atleast two different components (e.g., a compound disclosed herein and anacid), wherein each of the components is independently an atom, ion, ormolecule. In certain embodiments, none of the components is a solvent.In certain embodiments, at least one of the components is a solvent. Aco-crystal of a compound disclosed herein and an acid is different froma salt formed from a compound disclosed herein and the acid. In thesalt, a compound disclosed herein is complexed with the acid in a waythat proton transfer (e.g., a complete proton transfer) from the acid toa compound disclosed herein easily occurs at room temperature. In theco-crystal, however, a compound disclosed herein is complexed with theacid in a way that proton transfer from the acid to a compound disclosedherein does not easily occur at room temperature. In certainembodiments, in the co-crystal, there is no proton transfer from theacid to a compound disclosed herein. In certain embodiments, in theco-crystal, there is partial proton transfer from the acid to a compounddisclosed herein. Co-crystals may be useful to improve the properties(e.g., solubility, stability, and ease of formulation) of a compounddisclosed herein.

The term “polymorphs” refers to a crystalline form of a compound (or asalt, hydrate, or solvate thereof) in a particular crystal packingarrangement. All polymorphs have the same elemental composition.Different crystalline forms usually have different X-ray diffractionpatterns, infrared spectra, melting points, density, hardness, crystalshape, optical and electrical properties, stability, and solubility.Recrystallization solvent, rate of crystallization, storage temperature,and other factors may cause one crystal form to dominate. Variouspolymorphs of a compound can be prepared by crystallization underdifferent conditions.

The term “prodrugs” refer to compounds, including derivatives of thecompounds of Formula (I), which have cleavable groups and become bysolvolysis or under physiological conditions the compounds of Formula(I) which are pharmaceutically active in vivo. Such examples include,but are not limited to, ester derivatives and the like. Otherderivatives of the compounds of this disclosure have activity in boththeir acid and acid derivative forms, but in the acid sensitive formoften offers advantages of solubility, tissue compatibility, or delayedrelease in the mammalian organism (see, Bundgard, H., Design ofProdrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985). Prodrugs includeacid derivatives well known to practitioners of the art, such as, forexample, esters prepared by reaction of the parent acid with a suitablealcohol, or amides prepared by reaction of the parent acid compound witha substituted or unsubstituted amine, or acid anhydrides, or mixedanhydrides. Simple aliphatic or aromatic esters, amides, and anhydridesderived from acidic groups pendant on the compounds of this disclosureare particular prodrugs. In some cases it is desirable to prepare doubleester type prodrugs such as (acyloxy)alkyl esters or((alkoxycarbonyl)oxy)alkylesters. C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈alkynyl, aryl, C₆-C₁₂ substituted aryl, and C₆-C₁₂ arylalkyl esters ofthe compounds of Formula (I) may be preferred.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (i.e., a male or female of any age group, e.g., apediatric subject (e.g., infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult, or senior adult)) and/or othernon-human animals, for example, mammals (e.g., primates (e.g.,cynomolgus monkeys, rhesus monkeys); commercially relevant mammals, suchas cattle, pigs, horses, sheep, goats, cats, and/or dogs) and birds(e.g., commercially relevant birds such as chickens, ducks, geese,and/or turkeys). In certain embodiments, the animal is a mammal. Theanimal may be a male or female at any stage of development. A non-humananimal may be a transgenic animal. A subject who is resistant totreatment with a BTK inhibitor is one who shows no or minimal responseto the treatment. In some embodiments, response to a treatment ismeasured by reduction in tumor cells or tumor cell killing. In someembodiments, response to a treatment is measured by changes in symptomsof the disease, condition or malignancy (e.g., a proliferative disease).It has been discovered that the compounds that block ATP binding to HCKas described herein are able to cause tumor cell killing even in cellsthat are derived from subjects who are resistant to a BTK inhibitortreatment.

The term “biological sample” refers to any sample including tissuesamples (such as tissue sections and needle biopsies of a tissue); cellsamples (e.g., cytological smears (such as Pap or blood smears) orsamples of cells obtained by microdissection); samples of wholeorganisms (such as samples of yeasts or bacteria); or cell fractions,fragments, or organelles (such as obtained by lysing cells andseparating the components thereof by centrifugation or otherwise). Otherexamples of biological samples include blood, serum, urine, semen, fecalmatter, cerebrospinal fluid, interstitial fluid, mucous, tears, sweat,pus, biopsied tissue (e.g., obtained by a surgical biopsy or needlebiopsy), nipple aspirates, milk, vaginal fluid, saliva, swabs (such asbuccal swabs), or any material containing biomolecules that is derivedfrom a first biological sample.

The terms “administer,” “administering,” or “administration,” refers toimplanting, absorbing, ingesting, injecting, inhaling, applying, orotherwise introducing a compound, or a pharmaceutical compositionthereof, to a subject in need thereof, a biological sample, or a cell.

The terms “treatment,” “treat,” and “treating” refer to reversing,alleviating, delaying the onset of, or inhibiting the progress of a“pathological condition” (e.g., a disease, disorder, or condition, orone or more signs or symptoms thereof) described herein. In someembodiments, treatment may be administered after one or more signs orsymptoms have developed or have been observed. In other embodiments,treatment may be administered in the absence of signs or symptoms of thedisease or condition. For example, treatment may be administered to asusceptible individual prior to the onset of symptoms (e.g., in light ofa history of symptoms and/or in light of genetic or other susceptibilityfactors). Treatment may also be continued after symptoms have resolved,for example, to delay or prevent recurrence.

The terms “condition,” “disease,” and “disorder” are usedinterchangeably.

An “effective amount” of a compound of Formula (I) refers to an amountsufficient to elicit the desired biological response, i.e., treating thecondition. As will be appreciated by those of ordinary skill in thisart, the effective amount of a compound of Formula (I) may varydepending on such factors as the desired biological endpoint, thepharmacokinetics of the compound, the condition being treated, the modeof administration, and the age and health of the subject. An effectiveamount encompasses therapeutic and prophylactic treatment. For example,in treating cancer, an effective amount of a compound may reduce thetumor burden or stop the growth or spread of a tumor.

A “therapeutically effective amount” of a compound of Formula (I) is anamount sufficient to provide a therapeutic benefit in the treatment of acondition or to delay or minimize one or more symptoms associated withthe condition. A therapeutically effective amount of a compound means anamount of therapeutic agent, alone or in combination with othertherapies, which provides a therapeutic benefit in the treatment of thecondition. The term “therapeutically effective amount” can encompass anamount that improves overall therapy, reduces or avoids symptoms orcauses of the condition, or enhances the therapeutic efficacy of anothertherapeutic agent.

A “proliferative disease” refers to a disease that occurs due toabnormal growth or extension by the multiplication of cells (Walker,Cambridge Dictionary of Biology; Cambridge University Press: Cambridge,UK, 1990). A proliferative disease may be associated with: 1) thepathological proliferation of normally quiescent cells; 2) thepathological migration of cells from their normal location (e.g.,metastasis of neoplastic cells); 3) the pathological expression ofproteolytic enzymes such as the matrix metalloproteinases (e.g.,collagenases, gelatinases, and elastases); or 4) the pathologicalangiogenesis as in proliferative retinopathy and tumor metastasis.Exemplary proliferative diseases include cancers (i.e., “malignantneoplasms”), benign neoplasms, angiogenesis, inflammatory diseases, autoinflammatory diseases, and autoimmune diseases.

The terms “neoplasm” and “tumor” are used interchangeably and refer toan abnormal mass of tissue wherein the growth of the mass surpasses andis not coordinated with the growth of a normal tissue. A neoplasm ortumor may be “benign” or “malignant,” depending on the followingcharacteristics: degree of cellular differentiation (includingmorphology and functionality), rate of growth, local invasion, andmetastasis. A “benign neoplasm” is generally well differentiated, hascharacteristically slower growth than a malignant neoplasm, and remainslocalized to the site of origin. In addition, a benign neoplasm does nothave the capacity to infiltrate, invade, or metastasize to distantsites.

Exemplary benign neoplasms include, but are not limited to, lipoma,chondroma, adenomas, acrochordon, senile angiomas, seborrheic keratoses,lentigos, and sebaceous hyperplasias. In some cases, certain “benign”tumors may later give rise to malignant neoplasms, which may result fromadditional genetic changes in a subpopulation of the tumor's neoplasticcells, and these tumors are referred to as “pre-malignant neoplasms.” Anexemplary pre-malignant neoplasm is a teratoma. In contrast, a“malignant neoplasm” is generally poorly differentiated (anaplasia) andhas characteristically rapid growth accompanied by progressiveinfiltration, invasion, and destruction of the surrounding tissue.Furthermore, a malignant neoplasm generally has the capacity tometastasize to distant sites.

The term “metastasis,” “metastatic,” or “metastasize” refers to thespread or migration of cancerous cells from a primary or original tumorto another organ or tissue and is typically identifiable by the presenceof a “secondary tumor” or “secondary cell mass” of the tissue type ofthe primary or original tumor and not of that of the organ or tissue inwhich the secondary (metastatic) tumor is located. For example, aprostate cancer that has migrated to bone is said to be metastasizedprostate cancer and includes cancerous prostate cancer cells growing inbone tissue.

The term “cancer” refers to a malignant neoplasm (Stedman's MedicalDictionary, 25th ed.; Hensyl ed.; Williams & Wilkins: Philadelphia,1990). Exemplary cancers include, but are not limited to, acousticneuroma; adenocarcinoma; adrenal gland cancer; anal cancer; angiosarcoma(e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma);appendix cancer; benign monoclonal gammopathy; biliary cancer (e.g.,cholangiocarcinoma); bladder cancer; breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast); brain cancer (e.g., meningioma,glioblastomas, glioma (e.g., astrocytoma, oligodendroglioma),medulloblastoma); bronchus cancer; carcinoid tumor; cervical cancer(e.g., cervical adenocarcinoma); choriocarcinoma; chordoma;craniopharyngioma; colorectal cancer (e.g., colon cancer, rectal cancer,colorectal adenocarcinoma); connective tissue cancer; epithelialcarcinoma; ependymoma; endotheliosarcoma (e.g., Kaposi's sarcoma,multiple idiopathic hemorrhagic sarcoma); endometrial cancer (e.g.,uterine cancer, uterine sarcoma); esophageal cancer (e.g.,adenocarcinoma of the esophagus, Barrett's adenocarcinoma); Ewing'ssarcoma; eye cancer (e.g., intraocular melanoma, retinoblastoma);familiar hypereosinophilia; gall bladder cancer; gastric cancer (e.g.,stomach adenocarcinoma); gastrointestinal stromal tumor (GIST); germcell cancer; head and neck cancer (e.g., head and neck squamous cellcarcinoma, oral cancer (e.g., oral squamous cell carcinoma), throatcancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngealcancer, oropharyngeal cancer)); hematopoietic cancers (e.g., leukemiasuch as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL),acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronicmyelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chroniclymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL)); lymphomasuch as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) andnon-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large celllymphoma (DLCL) (e.g., diffuse large B-cell lymphoma), follicularlymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas(e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma),primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacyticlymphoma (i.e., Waldenström's macroglobulinemia), hairy cell leukemia(HCL), immunoblastic large cell lymphoma, precursor B-lymphoblasticlymphoma and primary central nervous system (CNS) lymphoma; and T-cellNHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheralT-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma,extranodal natural killer T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, and anaplasticlarge cell lymphoma); a mixture of one or more leukemia/lymphoma asdescribed above; and multiple myeloma (MM)), heavy chain disease (e.g.,alpha chain disease, gamma chain disease, mu chain disease);hemangioblastoma; hypopharynx cancer; inflammatory myofibroblastictumors; immunocytic amyloidosis; kidney cancer (e.g., nephroblastomaa.k.a. Wilms' tumor, renal cell carcinoma); liver cancer (e.g.,hepatocellular cancer (HCC), malignant hepatoma); lung cancer (e.g.,bronchogenic carcinoma, small cell lung cancer (SCLC), non-small celllung cancer (NSCLC), adenocarcinoma of the lung); leiomyosarcoma (LMS);mastocytosis (e.g., systemic mastocytosis); muscle cancer;myelodysplastic syndrome (MDS); mesothelioma; myeloproliferativedisorder (MPD) (e.g., polycythemia vera (PV), essential thrombocytosis(ET), agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF),chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML),chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES));neuroblastoma; neurofibroma (e.g., neurofibromatosis (NF) type 1 or type2, schwannomatosis); neuroendocrine cancer (e.g., gastroenteropancreaticneuroendocrinetumor (GEP-NET), carcinoid tumor); osteosarcoma (e.g.,bone cancer); ovarian cancer (e.g., cystadenocarcinoma, ovarianembryonal carcinoma, ovarian adenocarcinoma); papillary adenocarcinoma;pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductalpapillary mucinous neoplasm (IPMN), Islet cell tumors); penile cancer(e.g., Paget's disease of the penis and scrotum); pinealoma; primitiveneuroectodermal tumor (PNT); plasma cell neoplasia; paraneoplasticsyndromes; intraepithelial neoplasms; prostate cancer (e.g., prostateadenocarcinoma); rectal cancer; rhabdomyosarcoma; salivary gland cancer;skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA),melanoma, basal cell carcinoma (BCC)); small bowel cancer (e.g.,appendix cancer); soft tissue sarcoma (e.g., malignant fibroushistiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor(MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma); sebaceous glandcarcinoma; small intestine cancer; sweat gland carcinoma; synovioma;testicular cancer (e.g., seminoma, testicular embryonal carcinoma);thyroid cancer (e.g., papillary carcinoma of the thyroid, papillarythyroid carcinoma (PTC), medullary thyroid cancer); urethral cancer;vaginal cancer; and vulvar cancer (e.g., Paget's disease of the vulva).

The term “angiogenesis” refers to the formation and the growth of newblood vessels. Normal angiogenesis occurs in the healthy body of asubject for healing wounds and for restoring blood flow to tissues afterinjury. The healthy body controls angiogenesis through a number ofmeans, e.g., angiogenesis-stimulating growth factors and angiogenesisinhibitors. Many disease states, such as cancer, diabetic blindness,age-related macular degeneration, rheumatoid arthritis, and psoriasis,are characterized by abnormal (i.e., increased or excessive)angiogenesis. Abnormal or pathological angiogenesis refers toangiogenesis greater than that in a normal body, especially angiogenesisin an adult not related to normal angiogenesis (e.g., menstruation orwound healing). Abnormal angiogenesis can provide new blood vessels thatfeed diseased tissues and/or destroy normal tissues, and in the case ofcancer, the new vessels can allow tumor cells to escape into thecirculation and lodge in other organs (tumor metastases). In certainembodiments, the angiogenesis is pathological angiogenesis.

A “protein” or “peptide” comprises a polymer of amino acid residueslinked together by peptide bonds. The term refers to proteins,polypeptides, and peptides of any size, structure, or function.Typically, a protein will be at least three amino acids long. A proteinmay refer to an individual protein or a collection of proteins. Proteinspreferably contain only natural amino acids, although non-natural aminoacids (i.e., compounds that do not occur in nature but that can beincorporated into a polypeptide chain) and/or amino acid analogs as areknown in the art may alternatively be employed. Also, one or more of theamino acids in a protein may be modified, for example, by the additionof a chemical entity such as a carbohydrate group, a hydroxyl group, aphosphate group, a farnesyl group, an isofarnesyl group, a fatty acidgroup, a linker for conjugation or functionalization, or othermodification.

A protein may also be a single molecule or may be a multi-molecularcomplex. A protein may be a fragment of a naturally occurring protein orpeptide. A protein may be naturally occurring, recombinant, orsynthetic, or any combination of these.

The term “kinase” refers to any enzyme that catalyzes the addition ofphosphate groups to an amino acid residue of a substrate (e.g., aprotein or nucleoside). For example, a serine kinase catalyzes theaddition of a phosphate group to serine residue in a protein. In certainembodiments, the kinase is a tyrosine kinase. Examples of kinasesinclude, but are not limited to, a Janus kinase (e.g., SRC familykinases (e.g., HCK, LYN, BLK, FRK), Tec family kinases (e.g., BTK),Janus kinase 1 (JAK1), Janus kinase 2 (JAK2), Janus kinase 3 (JAK3),tyrosine kinase 2 (TYK2)), a CMGC kinase (e.g., a cyclin-dependentkinase (CDK, e.g., CDK1, CDK2, CDK2, CDK4, CDK5, CDK7, CDK8, CDK9,CDK10, CDK11, CDK12, CDK13, CDK14, CDK16, CDK20), a mitogen-activatedprotein kinase (MAPK, e.g., MAPK1, MAPK3, MAPK4, MAPK6, MAPK7, MAPK8,MAPK9, MAPK10, MAPK11, MAPK12, MAPK13, MAPK14, MAPK15), a glycogensynthase kinase 3 (GSK3, e.g., GSK3α, GSK3β), or a CDC-like kinase (CLK,e.g., CLK1, CLK2, CLK3, CLK4)), an AGC kinase (e.g., protein kinase A(PKA), protein kinase C (PKC), protein kinase G (PKG)), aCa²⁺/calmodulin-dependent protein kinase (CaM kinase, e.g., aspecialized CaM kinase, a multifunctional CaM kinase), a casein kinase 1(CK1, e.g., CK1alpha, CK1beta 1, CK1gamma 1, CK1gamma 2, CK1gamma 3,CK1delta, CK1epsilon), a STE kinase (e.g., a homolog of yeast Sterile 7,Sterile 11, or Sterile 20 kinase), a tyrosine kinase (TK, e.g., areceptor tyrosine kinase (RTK), a non-receptor tyrosine kinase (nRTK)),and a tyrosine-kinase-like kinase (TKL, e.g., a mixed lineage kinase(MLK), RAF, a serine threonine kinase receptor (STKR), a leucine richrepeat kinase (LRRK), a LIM domain kinase (LIMK), a testis expressedserine kinase (TESK), an IL1 receptor associated kinase (IRAK), areceptor interacting protein kinase (RIPK)).

HCK is a member of the src-family of protein tyrosine kinases, and isaberrantly up-regulated in WM cells. In myeloma cells, HCK is activatedby interleukin 6 (IL6) through the IL6 co-receptor IL6ST (GP130).

Bruton's tyrosine kinase (BTK) is a member of the src-related BTK/Tecfamily of cytoplasmic tyrosine kinases, is required for B cell receptorsignaling, plays a key role in B-cell maturation, and exhibits increasedactivation in a number of B-cell malignancies.

LYN proto-oncogene (LYN) is a member of the src-family of proteintyrosine kinases, plays an important role in the regulation of B-celldifferentiation, proliferation, survival, and apoptosis, is importantfor immune self-tolerance, and acts downstream of several immunereceptors, including the B-cell receptor (BCR). Without wishing to bebound by any particular theory, BCR signaling is thought to be involvedin pro-growth and survival signaling in MYD88 mutated disease, as wellas being involved in non-MYD88 mutated disease. For example, BCRsignaling is thought to be active in Waldenström's macroglobulinemia,ABC subtype of diffuse large B-cell lymphoma, and chronic lymphocyticleukemia.

Proto-oncogene tyrosine-protein kinase SRC (SRC) is a protein tyrosinekinase, plays a central role in the regulation of a variety ofbiological processes, such as cell proliferation, migration, adhesion,and survival in solid tumors, and is overexpressed in Waldenström'smacroglobulinemia.

As used herein “inhibition”, “inhibiting”, “inhibit,” and “inhibitor”,and the like, refer to the ability of a compound to reduce, slow, halt,block, or prevent activity of a particular biological process (e.g., akinase (e.g., SFK (e.g., HCK, LYN, BLK, FRK), Tec family kinases (e.g.,BTK)) in a cell relative to vehicle.

The terms “block” or “blocking” refer to the ability of a compound toprevent a biological interaction (e.g., binding) in a cell relative to anegative control, e.g., vehicle. For example, a compound can block ATPfrom binding to the ATP binding pocket of a kinase.

Such blocking may occur by direct binding of the compound to the ATPbinding pocket itself, or indirect blocking. In some embodiments, theterm refers to a reduction in the level of binding of ATP to a kinase(e.g., BTK, HCK, LYN, BLK, FRK, SRC) to a level that is statisticallysignificantly lower than an initial level, which may, for example, be abaseline level of ATP binding. In some embodiments, the term refers to areduction in the level of ATP binding to a kinase (e.g., BTK, HCK, LYN,BLK, FRK, SRC) to a level that is less than 75%, less than 50%, lessthan 40%, less than 30%, less than 25%, less than 20%, less than 10%,less than 9%, less than 8%, less than 7%, less than 6%, less than 5%,less than 4%, less than 3%, less than 2%, less than 1%, less than 0.5%,less than 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001%of an initial level, which may, for example, be a baseline level of ATPbinding. In some embodiments, blocking ATP binding leads to a reductionin the level of enzyme activity (e.g., BTK, HCK, LYN, BLK, FRK, SRCactivity) to a level that is less than 75%, less than 50%, less than40%, less than 30%, less than 25%, less than 20%, less than 10%, lessthan 9%, less than 8%, less than 7%, less than 6%, less than 5%, lessthan 4%, less than 3%, less than 2%, less than 1%, less than 0.5%, lessthan 0.1%, less than 0.01%, less than 0.001%, or less than 0.0001% of aninitial level, which may, for example, be a baseline level of enzymeactivity.

When a compound or pharmaceutical composition is referred to as“selectively,” “specifically,” or “competitively” binding a firstprotein, the compound binds the first protein, e.g., BTK, HCK, LYN, BLK,FRK, or SRC, with a higher binding affinity (e.g., not less than about2-fold, not less than about 5-fold, not less than about 10-fold, notless than about 30-fold, not less than about 100-fold, not less thanabout 1,000-fold, or not less than about 10,000-fold) than binding asecond protein that is different from the first protein, e.g., BTK. Insome embodiments, a compound blocks ATP binding to a first protein,e.g., HCK, LYN, BLK, FRK or SRC, at a lower concentration (e.g., notless than about 2-fold, not less than about 5-fold, not less than about10-fold, not less than about 30-fold, not less than about 100-fold, notless than about 1,000-fold, or not less than about 10,000-fold) than itblocks ATP binding a second protein that is different from the firstprotein, e.g., BTK.

Compounds which selectively block ATP binding to a kinase (e.g., BTK,HCK, LYN) provided herein can be identified and/or characterized bymethods known in the art. Methods include purified enzyme and cell basedbiochemical and binding assays such as an HCK gatekeeper mutant rescueassay, an in vitro kinase assay, e.g., using HCK gatekeeper mutatedkinase, competitive binding assays using KiNativ technology or biotintagged inhibitors, e.g., HCK inhibitors. Suitable assays for determiningselective inhibition of HCK by a compound include, but are not limitedto, Life Technology Z-Lyte activity assays (e.g., including HCKgatekeeper mutants and GK+6 mutants); DiscoverX KINOMEscan® bindingassays; MRC radioactivity assays; ACD Ba/F3 viability assays (e.g.,including HCK gatekeeper mutants and GK+6 mutants); Yeast hybridproliferation assays; Protein thermostability assays; and cancer cellswith HCK gatekeeper mutants or GK+6 mutants proliferation-rescue assays.Such assays can also be used to determine selective inhibition of LYNand/or SRC by a compound.

The term “MYD88 mutation” means any change or difference in the nucleicacid or protein sequence of MYD88 as compared to the wild type sequencethat results in the activation of MYD88 which leads to the activation ofNF-1B. Mutations include, but are not limited to, nonsense mutations,missense mutations, frameshift mutations, rearrangement mutations,insertion mutations, and deletion mutations. In some embodiments, themutation is a somatic mutation at position 38182641 in chromosome 3p22.2which results in a single nucleotide change from T→C in the myeloiddifferentiation primary response (MYD88) gene, and a predictednon-synonymous change at amino acid position 265 from leucine to proline(L265P). In some embodiments, the mutation is another activatingmutation in MYD88, such as V217F, W218R, I220T, S222R, M232T, S243N,T294P. Signaling studies show that SU-DHL-2 lymphoma cells that expressthe serine to arginine mutation at amino acid position 222 also haveupregulated HCK (Yang et al., Blood 2016). In some embodiments, Sangersequencing, whole exome or whole genome sequencing can be used toidentify somatic mutations in MYD88.

The term “MYD88 mutated disease” or “disease associated with mutatedMYD88” means any disease in a subject that is related to a change ordifference in the nucleic acid or protein sequence of MYD88 as comparedto the wild type sequence that results in the activation of MYD88 whichleads to the activation of NF-1B. In some embodiments, mutated MYD88disease is associated with a cancer (e.g., breast cancer, colon cancer,testicular cancer, CNS cancer, stomach cancer, lymphoma (e.g., B-celllymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgM secretinglymphoplasmacytic lymphoma (i.e., Waldenström's Macroglobulinemia),non-IgM secreting lymphoplasmacytic lymphoma)), diffuse large B-celllymphoma (e.g., activated B-cell-like (ABC)-DLBCL, germinal centerB-cell-like (GBC)-DLBCL), follicular lymphoma, marginal zone B-celllymphoma, small lymphocytic lymphoma, mantle cell lymphoma), andleukemia (e.g., chronic lymphocytic leukemia (CLL), acute lymphoblasticleukemia, myelogenous leukemia (e.g., chronic myelogenous leukemia,acute myelogenous leukemia))))). In some embodiments, mutated MYD88 isassociated with susceptibility to an infectious disease. In someembodiments, mutated MYD88 is associated with susceptibility to anautoimmune disease.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which constitute a part of thisspecification, illustrate several embodiments of the disclosure andtogether with the description, serve to explain the principles of thedisclosure.

FIG. 1 shows pro-survival signaling driven by mutated MYD88 and thepossible kinases targeted by the compounds of Formula (I).

FIG. 2 shows a dose-response curves for Compound I-4, as measured byCellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WMcells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and multiple myelomacells.

FIG. 3 shows a dose-response curves for Compound I-1, as measured byCellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WMcells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and multiple myelomacells.

FIG. 4 shows the drug dose-response for ibrutinib and Compound I-4 inBTK WT or BTK C481S expressing lentiviral vector transduced TMD8ABC-DLBCL or BCWM.1 WM cells.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION

Kinases are implicated in a wide variety of diseases, e.g.,proliferative diseases. Provided herein are compounds of Formula (I),and pharmaceutically acceptable salts, solvates, hydrates, polymorphs,co-crystals, tautomers, stereoisomers, isotopically labeled derivatives,and prodrugs thereof. The provided compounds may be kinase inhibitors.In certain embodiments, the kinase being targeted is an SRC familykinase (e.g., HCK, LYN, BLK, FRK). In certain embodiments, the kinasebeing targeted is a Tec family kinase (e.g., BTK). Also provided arepharmaceutical compositions and kits comprising the provided compounds.Further provided are methods of using the compounds, pharmaceuticalcompositions, and kits for treating a disease (e.g., proliferativedisease) in a subject in need thereof. In certain embodiments, thedisease is a proliferative disease (e.g., an IgM gammopathy (e.g., anIgM Monoclonal gammopathy of undetermined significance (MGUS), amyloidlight chain (AL) amyloidosis), mastocytosis (e.g., systemicmastocytosis), cancer (e.g., breast cancer, colon cancer, testicularcancer, CNS cancer, stomach cancer, lymphoma (e.g., B-cell lymphoma(e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacyticlymphoma (i.e., Waldenström's Macroglobulinemia), non-IgM secretinglymphoplasmacytic lymphoma)), diffuse large B-cell lymphoma (e.g.,activated B-cell-like (ABC)-DLBCL, germinal center B-cell-like(GBC)-DLBCL), follicular lymphoma, marginal zone B-cell lymphoma, smalllymphocytic lymphoma, mantle cell lymphoma), myeloma (e.g., IgM myelomas(e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocyticleukemia (CLL), acute lymphoblastic leukemia, myelogenous leukemia(e.g., chronic myelogenous leukemia, acute myelogenous leukemia (e.g.,mast cell leukemia), myeloproliferative diseases (e.g., myelodysplasticsyndrome))))) Further provided are methods of using the providedcompounds, pharmaceutical compositions, and kits for inhibiting theactivity of a kinase in a subject in need thereof, in a biologicalsample, or in a cell.

Compounds

In one aspect of the present disclosure, provided herein are compoundsof Formula (I):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof, wherein:

R is optionally substituted alkynyl, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl,optionally substituted heteroaryl, —O-optionally substitutedcarbocyclyl, —CH₂—O-optionally substituted carbocyclyl, —O-optionallysubstituted aryl, —CH₂—O-optionally substituted aryl, —O-optionallysubstituted heteroaryl, or

—CH₂—O-optionally substituted heteroaryl;

R¹ is hydrogen, optionally substituted acyl, optionally substitutedalkyl, optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl, or anitrogen protecting group;

each occurrence of R² is independently hydrogen, halogen, optionallysubstituted acyl, optionally substituted alkyl, optionally substitutedalkenyl, optionally substituted alkynyl, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, optionally substituted heteroaryl, —OR^(a), —N(R^(a))₂, or—SR^(a), wherein R^(a) is independently selected from hydrogen,optionally substituted acyl, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, an oxygenprotecting group when attached to an oxygen atom, and a sulfurprotecting group when attached to a sulfur atom;

each occurrence of R³ is independently hydrogen, halogen, optionallysubstituted acyl, optionally substituted alkyl, optionally substitutedalkenyl, optionally substituted alkynyl, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, optionally substituted heteroaryl, —OR^(b), —N(R^(b))₂, or—SR^(b), wherein R^(b) is independently selected from hydrogen,optionally substituted acyl, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, an oxygenprotecting group when attached to an oxygen atom, and a sulfurprotecting group when attached to a sulfur atom;

J is a bond or alkynylene;

m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, as valency permits;

n is 1, 2, 3, 4, 5, 6, 7, or 8, as valency permits;

A is an optionally substituted mono- or bicyclic-heteroaryl, oroptionally substituted bicyclic heterocyclyl; and

provided that when J is a bond, R is substituted phenyl, R¹ is methyl,R² is H, and R³ is H, then ring A is not of the formula:

Formula (I), as described herein, contains the substituent R. In certainembodiments, R is optionally substituted alkynyl. In certainembodiments, R is optionally substituted carbocyclyl. In certainembodiments, R is optionally substituted heterocyclyl. In certainembodiments, R is optionally substituted aryl. In certain embodiments, Ris optionally substituted phenyl. In certain embodiments, R is4-phenoxyphenyl. In certain embodiments, R is optionally substitutedheteroaryl. In certain embodiments, R is —O-optionally substitutedcarbocyclyl. In certain embodiments, R is —CH₂—O-optionally substitutedcarbocyclyl. In certain embodiments, R is —O-optionally substitutedaryl. In certain embodiments, R is —CH₂—O-optionally substituted aryl.In certain embodiments, R is —CH₂—O-optionally substituted phenyl. Incertain embodiments, R is —CH₂—O-unsubstituted phenyl. In certainembodiments, R is —O-optionally substituted heteroaryl. In certainembodiments, R is —CH₂—O-optionally substituted heteroaryl.

Formula (I) contains the substituent R¹. In certain embodiments, R¹ ishydrogen. In certain embodiments, R¹ is optionally substituted acyl. Incertain embodiments, R¹ is optionally substituted alkyl (e.g., Me, Et,Pr). In certain embodiments, R¹ is optionally substituted methyl. Incertain embodiments, R¹ is unsubstituted methyl. In certain embodiments,R¹ is optionally substituted alkenyl. In certain embodiments, R¹ isoptionally substituted alkynyl. In certain embodiments, R¹ is optionallysubstituted carbocyclyl. In certain embodiments, R¹ is optionallysubstituted heterocyclyl. In certain embodiments, R¹ is optionallysubstituted aryl. In certain embodiments, R¹ is optionally substitutedheteroaryl. In certain embodiments, R¹ is a nitrogen protecting group.

In certain embodiments, R¹ is methyl.

Formula (I) contains the substituents R². In certain embodiments, atleast one of R² is hydrogen. In certain embodiments, at least oneinstance of R² is halogen. In certain embodiments, at least one instanceof R² is optionally substituted acyl. In certain embodiments, at leastone instance of R² is optionally substituted alkyl. In certainembodiments, at least one instance of R² is optionally substitutedalkenyl. In certain embodiments, at least one instance of R² isoptionally substituted alkynyl. In certain embodiments, at least oneinstance of R² is optionally substituted carbocyclyl. In certainembodiments, at least one instance of R² is optionally substitutedheterocyclyl. In certain embodiments, at least one instance of R² isoptionally substituted aryl. In certain embodiments, at least oneinstance of R² is optionally substituted heteroaryl. In certainembodiments, at least one instance of R² is —OR^(a). In certainembodiments, at least one instance of R² is —N(R^(a))₂.

In certain embodiments, R² is H.

In certain embodiments, there are m instances of R². In certainembodiments, m is 1. In certain embodiments, m is 2. In certainembodiments, m is 3. In certain embodiments, m is 4. In certainembodiments, m is 5. In certain embodiments, m is 6. In certainembodiments, m is 7. In certain embodiments, m is 8. In certainembodiments, m is 9. In certain embodiments, m is 10.

In certain embodiments, R² is H; and m is 1.

Formula (I) contains the substituents R³. In certain embodiments, atleast one instance of R³ is hydrogen. In certain embodiments, at leastone instance of R³ is halogen. In certain embodiments, at least oneinstance of R³ is optionally substituted acyl. In certain embodiments,at least one instance of R³ is optionally substituted alkyl. In certainembodiments, at least one instance of R³ is optionally substitutedalkenyl. In certain embodiments, at least one instance of R³ isoptionally substituted alkynyl. In certain embodiments, at least oneinstance of R³ is optionally substituted carbocyclyl. In certainembodiments, at least one instance of R³ is optionally substitutedheterocyclyl. In certain embodiments, at least one instance of R³ isoptionally substituted aryl. In certain embodiments, at least oneinstance of R³ is optionally substituted heteroaryl. In certainembodiments, at least one instance of R³ is —OR^(b). In certainembodiments, at least one instance of R³ is —N(R^(b))₂. In certainembodiments, at least one instance of R³ is —SR^(b).

In certain embodiments, R³ is H.

In certain embodiments, there are n instances of R³. In certainembodiments, n is 1. In certain embodiments, n is 2. In certainembodiments, n is 3. In certain embodiments, n is 4. In certainembodiments, n is 5. In certain embodiments, n is 6. In certainembodiments, n is 7. In certain embodiments, n is 8.

In certain embodiments, R³ is H; and n is 1.

In certain embodiments, R² and R³ are the same. In certain embodiments,R² and R³ are different. In certain embodiments, R² is H; and R³ is H.

Formula (I) contains the substituent J. In certain embodiments, J is abond. In certain embodiments, J is alkynylene.

Formula (I) contains ring A. In certain embodiments, ring A is of theformula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, J is alkynyl; and ring A is of the formula:

In certain embodiments, J is alkynyl ring A is of the formula:

In certain embodiments, J is alkynyl and ring A is of the formula:

In certain embodiments, J is alkynyl ring A is of the formula:

In certain embodiments, J is alkynyl and ring A is of the formula:

In certain embodiments, Ring A comprises the substituent R⁴. In certainembodiments, each instance of R⁴ is the same. In certain embodiments,each instance of R⁴ is different. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is halogen. In certain embodiments,R⁴ is optionally substituted acyl. In certain embodiments, R⁴ isoptionally substituted alkyl (e.g., Me, Et, Pr). In certain embodiments,R⁴ is —CN. In certain embodiments, R⁴ is —SCN. In certain embodiments,R⁴ is —NO₂. In certain embodiments, R⁴ is —C(═O)R^(c). In certainembodiments, R⁴ is —C(═O)N(R^(c))₂. In certain embodiments, R⁴ is—OR^(c). In certain embodiments, R⁴ is —N(R^(c))₂. In certainembodiments, R⁴ is —C(═NR^(c))R^(c). In certain embodiments, R⁴ is—C(═NR^(c))OR^(c). In certain embodiments, R⁴ is —C(═NR^(c))N(R^(c))₂.In certain embodiments, R⁴ is —C(═O)OR^(c). In certain embodiments, R⁴is —NR^(c)C(═O)R^(c). In certain embodiments, R⁴ is —NR^(c)C(═O)OR^(c).In certain embodiments, R⁴ is —NR^(c)C(═O)N(R^(c))₂. In certainembodiments, R⁴ is —OC(═O)R^(c). In certain embodiments, R⁴ is—OC(═O)OR^(c). In certain embodiments, R⁴ is —OC(═O)N(R^(c))₂. Incertain embodiments, R⁴ is —SRC, wherein R^(C) is independently selectedfrom hydrogen, optionally substituted acyl, optionally substitutedalkyl, optionally substituted alkenyl, optionally substituted alkynyl,an oxygen protecting group when attached to an oxygen atom, and a sulfurprotecting group when attached to a sulfur atom.

In certain embodiments, R⁴ is H.

In certain embodiments, R⁴ is —C(═O)N(R^(c))₂. In certain embodiments,R⁴ is —C(═O)NH₂.

In certain embodiments, Ring A comprises the substituent R⁵. In certainembodiments, at R⁵ is hydrogen. In certain embodiments, R⁵ is halogen.In certain embodiments, R⁵ is optionally substituted acyl. In certainembodiments, R⁵ is optionally substituted alkyl (e.g., Me, Et, Pr). Incertain embodiments, R⁵ is —CN. In certain embodiments, R⁵ is —SCN. Incertain embodiments, R⁵ is —NO₂. In certain embodiments, R⁵ is—C(═O)R^(d). In certain embodiments, R⁵ is —C(═O)N(R^(d))₂. In certainembodiments, R⁵ is —OR^(d). In certain embodiments, R⁵ is —N(R^(d))₂. Incertain embodiments, R⁵ is —C(═NR^(d))R^(d). In certain embodiments, R⁵is —C(═NR^(d))OR^(d). In certain embodiments, R⁵ is—C(═NR^(d))N(R^(d))₂. In certain embodiments, R⁵ is —C(═O)OR^(d). Incertain embodiments, R⁵ is —NR^(d)C(═O)R^(d). In certain embodiments, R⁵is —NR^(d)C(═O)OR^(d). In certain embodiments, R⁵ is—NR^(d)C(═O)N(R^(d))₂. In certain embodiments, R⁵ is —OC(═O)R^(d). Incertain embodiments, R⁵ is —OC(═O)OR^(d). In certain embodiments, R⁵ is—OC(═O)N(R^(d))₂. In certain embodiments, R⁵ is —SR^(d), wherein R^(d)is independently selected from hydrogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, an oxygen protecting group when attached to anoxygen atom, and a sulfur protecting group when attached to a sulfuratom.

In certain embodiments, R⁵ is H. In certain embodiments, R⁵ is—N(R^(d))₂. In certain embodiments, R⁵ is —NH₂.

In certain embodiments, R⁴ is —C(═O)N(R^(d))₂; and R⁵ is —N(R^(d))₂. Incertain embodiments, R⁴ is —C(═O)NH₂; and R⁵ is —N(R^(d))₂. In certainembodiments, R⁴ is —C(═O)N(R^(c))₂; and R⁵ is —NH₂. In certainembodiments, R⁴ is —C(═O)NH₂; and R⁵ is —NH₂.

In certain embodiments, Ring A comprises the substituent R⁶. In certainembodiments, at R⁶ is hydrogen. In certain embodiments, R⁶ is halogen.In certain embodiments, R⁶ is optionally substituted acyl. In certainembodiments, R⁶ is optionally substituted alkyl (e.g., Me, Et, Pr). Incertain embodiments, R⁶ is —CN. In certain embodiments, R⁶ is —SCN. Incertain embodiments, R⁶ is —NO₂. In certain embodiments, R⁶ is—C(═O)R^(e). In certain embodiments, R⁶ is —C(═O)N(R^(e))₂. In certainembodiments, R⁶ is —OR^(d). In certain embodiments, R⁶ is —N(R^(e))₂. Incertain embodiments, R⁶ is —C(═NR^(e))R^(e). In certain embodiments, R⁶is —C(═NR^(e))OR^(e). In certain embodiments, R⁶ is—C(═NR^(e))N(R^(e))₂. In certain embodiments, R⁶ is —C(═O)OR^(e). Incertain embodiments, R⁶ is —NR^(e)C(═O)R^(e). In certain embodiments, R⁶is —NR^(e)C(═O)OR^(e). In certain embodiments, R⁶ is—NR^(e)C(═O)N(R^(e))₂. In certain embodiments, R⁶ is —OC(═O)R^(e). Incertain embodiments, R⁶ is —OC(═O)OR^(e). In certain embodiments, R⁶ is—OC(═O)N(R^(e))₂. In certain embodiments, R⁶ is —SR^(c), wherein R^(c)is independently selected from hydrogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, an oxygen protecting group when attached to anoxygen atom, and a sulfur protecting group when attached to a sulfuratom.

In certain embodiments, R⁶ is H.

In certain embodiments, ring A contains the substituent X¹. In certainembodiments, X¹═N—. In certain embodiments, X¹ is ═C(R^(ff))—. Incertain embodiments, R^(f) is hydrogen, halogen, or substituted orunsubstituted C₁₋₆ alkyl. In certain embodiments, X¹ is ═C(H)—.

In certain embodiments, ring A contains the substituent X². In certainembodiments, X²═N—. In certain embodiments, X² is ═C(R^(g))—. In certainembodiments, R^(g) is hydrogen, halogen, or substituted or unsubstitutedC₁₋₆ alkyl. In certain embodiments, X² is ═C(H)—.

In certain embodiments, ring A contains the substituent X³. In certainembodiments, X³═N—. In certain embodiments, X³ is ═C(R^(g))—. In certainembodiments, R^(g) is hydrogen, halogen, or substituted or unsubstitutedC₁₋₆ alkyl. In certain embodiments, X² is ═C(H)—.

In certain embodiments, ring A contains the substituent X⁴. In certainembodiments, X⁴═N—. In certain embodiments, X⁴ is ═C(R^(ff))—. Incertain embodiments, R^(f) is hydrogen, halogen, or substituted orunsubstituted C₁₋₆ alkyl. In certain embodiments, X¹ is ═C(H)—.

In certain embodiments, ring A contains the substituent X⁴. In certainembodiments, X⁴ is ═N—. In certain embodiments, X⁴ is ═C(R¹)—. Incertain embodiments, X⁴ is ═C(H)—. In certain embodiments, R¹ ishydrogen, halogen, or substituted or unsubstituted C₁₋₆ alkyl (e.g., Me,Et, Pr).

In certain embodiments, X⁴ is ═N—; and R is —CH₂—O-optionallysubstituted aryl. In certain embodiments, X⁴ is ═C(R^(ff))—; and R is—CH₂—O-optionally substituted aryl. In certain embodiments, X⁴ is═C(H)—; and R is —CH₂—O-optionally substituted aryl. In certainembodiments, X⁴ is ═N—; and R is —CH₂—O-unsubstituted phenyl. In certainembodiments, X⁴ is ═C(R¹)—; and R is —CH₂—O-unsubstituted phenyl. Incertain embodiments, X⁴ is ═C(H)— and R is —CH₂—O-unsubstituted phenyl.

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is of the formula:

In certain embodiments, ring A is not of the formula:

In certain embodiments, ring A is not of the formula:

In certain embodiments, ring A is not of the formula:

In certain embodiments, ring A is not of the formula:

In certain embodiments, ring A is not of the formula:

In certain embodiments, J is a bond; and ring A is not of the formula:

In certain embodiments, J is a bond; and ring A is not of the formula:

In certain embodiments, J is a bond; and ring A is not of the formula:

In certain embodiments, J is a bond; and ring A is not of the formula:

In certain embodiments, a compound of Formula (I) is of the Formula(II):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the Formula(III):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the Formula(IV):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the Formula (V):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, the compound of Formula (V) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, the compound of Formula (V) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the Formula(VI):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof.

In certain embodiments, a compound of Formula (I) is a compound, orpharmaceutically acceptable salt thereof, of the formula:

# Structure I-1

I-2

I-3

I-4

I-5

I-6

I-7

I-8

I-9

I-10

Pharmaceutical Compositions, Administration, and Kits

The present disclosure also provides pharmaceutical compositionscomprising a compound described herein and optionally a pharmaceuticallyacceptable excipient. In certain embodiments, the pharmaceuticalcomposition further comprises an additional pharmaceutical agent.

In certain embodiments, the compound described herein is provided in aneffective (e.g., effective for inhibiting a kinase, such as BTK, HCK,LYN, BLK, FRK) amount in the pharmaceutical composition. In certainembodiments, the effective amount is a therapeutically effective amount.In certain embodiments, a therapeutically effective amount is an amounteffective for inhibiting a kinase. In certain embodiments, atherapeutically effective amount is an amount effective for treating adisease (e.g., a disease associated with aberrant activity of a kinase(e.g., proliferative disease)). In certain embodiments, atherapeutically effective amount is an amount effective for inhibitingthe activity of a kinase and treating a disease (e.g., a diseaseassociated with aberrant activity of a kinase (e.g., proliferativedisease (e.g., cancer (e.g., breast cancer, colon cancer, testicularcancer, CNS cancer, stomach cancer, lymphoma (e.g., B-cell lymphoma(e.g., lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacyticlymphoma (i.e., Waldenström's Macroglobulinemia), non-IgM secretinglymphoplasmacytic lymphoma)), diffuse large B-cell lymphoma (e.g.,activated B-cell-like (ABC)-DLBCL, germinal center B-cell-like(GBC)-DLBCL), follicular lymphoma, marginal zone B-cell lymphoma, smalllymphocytic lymphoma, mantle cell lymphoma), and leukemia (e.g., chroniclymphocytic leukemia (CLL), acute lymphoblastic leukemia, myelogenousleukemia (e.g., chronic myelogenous leukemia, acute myelogenousleukemia))))).

In certain embodiments, the effective amount is an amount effective forinhibiting the activity of a kinase by at least 10%, at least 20%, atleast 30%, at least 40%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, at least 95%, or at least 98%. In certainembodiments, the effective amount is an amount effective for inhibitingthe activity of a kinase by not more than 10%, not more than 20%, notmore than 30%, not more than 40%, not more than 50%, not more than 60%,not more than 70%, not more than 80%, not more than 90%, not more than95%, or not more than 98%.

In certain embodiments, the subject is an animal. The animal may be ofeither sex and may be at any stage of development. In certainembodiments, the subject described herein is a human (e.g., an adult,juvenile, or child). In certain embodiments, the subject is a non-humananimal. In certain embodiments, the subject is a mammal. In certainembodiments, the subject is a non-human mammal. In certain embodiments,the subject is a domesticated animal, such as a dog, cat, cow, pig,horse, sheep, or goat. In certain embodiments, the subject is a dog. Incertain embodiments, the subject is a companion animal, such as a dog orcat. In certain embodiments, the subject is a livestock animal, such asa cow, pig, horse, sheep, or goat. In certain embodiments, the subjectis a zoo animal. In another embodiment, the subject is a researchanimal, such as a rodent (e.g., mouse, rat), dog, pig, or non-humanprimate. In certain embodiments, the subject is a genetically engineeredanimal. In certain embodiments, the subject is a transgenic animal(e.g., transgenic mice, transgenic pigs). In certain embodiments, thesubject is a fish or reptile.

In certain embodiments, the biological sample or cell (e.g., thebiological sample or cell being contacted with a compound orpharmaceutical composition described herein) is in vitro. In certainembodiments, the biological sample or cell is in vivo or ex vivo. Incertain embodiments, the cell is a malignant cell or premalignant cell.

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include bringing the compound described herein (i.e., the“active ingredient”) into association with a carrier or excipient,and/or one or more other accessory ingredients, and then, if necessaryand/or desirable, shaping, and/or packaging the product into a desiredsingle- or multi-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.A “unit dose” is a discrete amount of the pharmaceutical compositioncomprising a predetermined amount of the active ingredient. The amountof the active ingredient is generally equal to the dosage of the activeingredient which would be administered to a subject and/or a convenientfraction of such a dosage, such as one-half or one-third of such adosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition described herein will vary, depending uponthe identity, size, and/or condition of the subject treated and furtherdepending upon the route by which the composition is to be administered.The composition may comprise between 0.1% and 100% (w/w) activeingredient.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

Exemplary diluents include calcium carbonate, sodium carbonate, calciumphosphate, dicalcium phosphate, calcium sulfate, calcium hydrogenphosphate, sodium phosphate lactose, sucrose, cellulose,microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodiumchloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

Exemplary granulating and/or dispersing agents include potato starch,corn starch, tapioca starch, sodium starch glycolate, clays, alginicacid, guar gum, citrus pulp, agar, bentonite, cellulose, and woodproducts, natural sponge, cation-exchange resins, calcium carbonate,silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone)(crospovidone), sodium carboxymethyl starch (sodium starch glycolate),carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose(croscarmellose), methylcellulose, pregelatinized starch (starch 1500),microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate,quaternary ammonium compounds, and mixtures thereof.

Exemplary surface active agents and/or emulsifiers include naturalemulsifiers (e.g., acacia, agar, alginic acid, sodium alginate,tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk,casein, wool fat, cholesterol, wax, and lecithin), colloidal clays(e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminumsilicate)), long chain amino acid derivatives, high molecular weightalcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetinmonostearate, ethylene glycol distearate, glyceryl monostearate, andpropylene glycol monostearate, polyvinyl alcohol), carbomers (e.g.,carboxy polymethylene, polyacrylic acid, acrylic acid polymer, andcarboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,carboxymethylcellulose sodium, powdered cellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylenesorbitan monolaurate (Tween® 20), polyoxyethylene sorbitan (Tween® 60),polyoxyethylene sorbitan monooleate (Tween® 80), sorbitan monopalmitate(Span® 40), sorbitan monostearate (Span® 60), sorbitan tristearate(Span® 65), glyceryl monooleate, sorbitan monooleate (Span® 80),polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj® 45),polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil,polyoxymethylene stearate, and Solutol*), sucrose fatty acid esters,polyethylene glycol fatty acid esters (e.g., Cremophor®),polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij® 30)),poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamineoleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium lauryl sulfate, Pluronic® F-68, poloxamer P-188,cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride,docusate sodium, and/or mixtures thereof.

Exemplary binding agents include starch (e.g., cornstarch and starchpaste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin,molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums(e.g., acacia, sodium alginate, extract of Irish moss, panwar gum,ghatti gum, mucilage of isapol husks, carboxymethylcellulose,methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate(Veegum©), and larch arabogalactan), alginates, polyethylene oxide,polyethylene glycol, inorganic calcium salts, silicic acid,polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

Exemplary preservatives include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, antiprotozoanpreservatives, alcohol preservatives, acidic preservatives, and otherpreservatives. In certain embodiments, the preservative is anantioxidant. In other embodiments, the preservative is a chelatingagent.

Exemplary antioxidants include alpha tocopherol, ascorbic acid, ascorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, potassium metabisulfite, propionic acid, propylgallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, andsodium sulfite.

Exemplary chelating agents include ethylenediaminetetraacetic acid(EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodiumedetate, trisodium edetate, calcium disodium edetate, dipotassiumedetate, and the like), citric acid and salts and hydrates thereof(e.g., citric acid monohydrate), fumaric acid and salts and hydratesthereof, malic acid and salts and hydrates thereof, phosphoric acid andsalts and hydrates thereof, and tartaric acid and salts and hydratesthereof. Exemplary antimicrobial preservatives include benzalkoniumchloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide,cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol,chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,propylene glycol, and thimerosal.

Exemplary antifungal preservatives include butyl paraben, methylparaben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoicacid, potassium benzoate, potassium sorbate, sodium benzoate, sodiumpropionate, and sorbic acid.

Exemplary alcohol preservatives include ethanol, polyethylene glycol,phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate,and phenylethyl alcohol.

Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E,beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbicacid, sorbic acid, and phytic acid.

Other preservatives include tocopherol, tocopherol acetate, deteroximemesylate, cetrimide, butylated hydroxyanisol (BHA), butylatedhydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS),sodium lauryl ether sulfate (SLES), sodium bisulfite, sodiummetabisulfite, potassium sulfite, potassium metabisulfite, Glydant®Plus, Phenonip®, methylparaben, Germall® 115, Germaben® II, Neolone®,Kathon®, and Euxyl®.

Exemplary buffering agents include citrate buffer solutions, acetatebuffer solutions, phosphate buffer solutions, ammonium chloride, calciumcarbonate, calcium chloride, calcium citrate, calcium glubionate,calcium gluceptate, calcium gluconate, D-gluconic acid, calciumglycerophosphate, calcium lactate, propanoic acid, calcium levulinate,pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasiccalcium phosphate, calcium hydroxide phosphate, potassium acetate,potassium chloride, potassium gluconate, potassium mixtures, dibasicpotassium phosphate, monobasic potassium phosphate, potassium phosphatemixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodiumcitrate, sodium lactate, dibasic sodium phosphate, monobasic sodiumphosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide,aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline,Ringer's solution, ethyl alcohol, and mixtures thereof.

Exemplary lubricating agents include magnesium stearate, calciumstearate, stearic acid, silica, talc, malt, glyceryl behanate,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate,sodium lauryl sulfate, and mixtures thereof.

Exemplary natural oils include almond, apricot kernel, avocado, babassu,bergamot, black current seed, borage, cade, camomile, canola, caraway,carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee,corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed,geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate,jojoba, kukui nut, lavandin, lavender, lemon, Litsea cubeba, macademianut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, andwheat germ oils. Exemplary synthetic oils include, but are not limitedto, butyl stearate, caprylic triglyceride, capric triglyceride,cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate,mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixturesthereof.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents. Incertain embodiments for parenteral administration, the conjugatesdescribed herein are mixed with solubilizing agents such as Cremophor®,alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins,polymers, and mixtures thereof.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension, or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, U.S.P., and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or di-glycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This can be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility.

The rate of absorption of the drug then depends upon its rate ofdissolution, which, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered drug form may be accomplished by dissolving or suspendingthe drug in an oil vehicle.

Compositions for rectal or vaginal administration are typicallysuppositories which can be prepared by mixing the conjugates describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol, or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active ingredient.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or (a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, (b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, (c) humectants such as glycerol, (d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, (e) solutionretarding agents such as paraffin, (f) absorption accelerators such asquaternary ammonium compounds, (g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolinand bentonite clay, and (i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets, and pills, thedosage form may include a buffering agent.

Solid compositions of a similar type can be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the art of pharmacology. Theymay optionally comprise opacifying agents and can be of a compositionthat they release the active ingredient(s) only, or preferentially, in acertain part of the intestinal tract, optionally, in a delayed manner.Examples of encapsulating compositions which can be used includepolymeric substances and waxes. Solid compositions of a similar type canbe employed as fillers in soft and hard-filled gelatin capsules usingsuch excipients as lactose or milk sugar as well as high molecularweight polyethylene glycols and the like.

The active ingredient can be in a micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings, and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active ingredient can be admixed with at least oneinert diluent such as sucrose, lactose, or starch. Such dosage forms maycomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may comprise bufferingagents. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of encapsulating agents which can be usedinclude polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of a compounddescribed herein may include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants, and/or patches. Generally, theactive ingredient is admixed under sterile conditions with apharmaceutically acceptable carrier or excipient and/or any neededpreservatives and/or buffers as can be required. Additionally, thepresent disclosure contemplates the use of transdermal patches, whichoften have the added advantage of providing controlled delivery of anactive ingredient to the body. Such dosage forms can be prepared, forexample, by dissolving and/or dispensing the active ingredient in theproper medium. Alternatively or additionally, the rate can be controlledby either providing a rate controlling membrane and/or by dispersing theactive ingredient in a polymer matrix and/or gel.

Suitable devices for use in delivering intradermal pharmaceuticalcompositions described herein include short needle devices. Intradermalcompositions can be administered by devices which limit the effectivepenetration length of a needle into the skin. Alternatively oradditionally, conventional syringes can be used in the classical mantouxmethod of intradermal administration. Jet injection devices whichdeliver liquid formulations to the dermis via a liquid jet injectorand/or via a needle which pierces the stratum corneum and produces a jetwhich reaches the dermis are suitable. Ballistic powder/particledelivery devices which use compressed gas to accelerate the compound inpowder form through the outer layers of the skin to the dermis aresuitable.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi-liquid preparations such as liniments,lotions, oil-in-water and/or water-in-oil emulsions such as creams,ointments, and/or pastes, and/or solutions and/or suspensions. Topicallyadministrable formulations may, for example, comprise from about 1% toabout 10% (w/w) active ingredient, although the concentration of theactive ingredient can be as high as the solubility limit of the activeingredient in the solvent. Formulations for topical administration mayfurther comprise one or more of the additional ingredients describedherein.

A pharmaceutical composition described herein can be prepared, packaged,and/or sold in a formulation suitable for pulmonary administration viathe buccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers, or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration using a device comprising a dry powderreservoir to which a stream of propellant can be directed to dispersethe powder and/or using a self-propelling solvent/powder dispensingcontainer such as a device comprising the active ingredient dissolvedand/or suspended in a low-boiling propellant in a sealed container. Suchpowders comprise particles wherein at least 98% of the particles byweight have a diameter greater than 0.5 nanometers and at least 95% ofthe particles by number have a diameter less than 7 nanometers.Alternatively, at least 95% of the particles by weight have a diametergreater than 1 nanometer and at least 90% of the particles by numberhave a diameter less than 6 nanometers. Dry powder compositions mayinclude a solid fine powder diluent such as sugar and are convenientlyprovided in a unit dose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions described herein formulated for pulmonarydelivery may provide the active ingredient in the form of droplets of asolution and/or suspension. Such formulations can be prepared, packaged,and/or sold as aqueous and/or dilute alcoholic solutions and/orsuspensions, optionally sterile, comprising the active ingredient, andmay conveniently be administered using any nebulization and/oratomization device. Such formulations may further comprise one or moreadditional ingredients including, but not limited to, a flavoring agentsuch as saccharin sodium, a volatile oil, a buffering agent, a surfaceactive agent, and/or a preservative such as methylhydroxybenzoate. Thedroplets provided by this route of administration may have an averagediameter in the range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition describedherein. Another formulation suitable for intranasal administration is acoarse powder comprising the active ingredient and having an averageparticle from about 0.2 to 500 micrometers. Such a formulation isadministered by rapid inhalation through the nasal passage from acontainer of the powder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) to as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein. A pharmaceutical composition described herein can beprepared, packaged, and/or sold in a formulation for buccaladministration. Such formulations may, for example, be in the form oftablets and/or lozenges made using conventional methods, and maycontain, for example, 0.1 to 20% (w/w) active ingredient, the balancecomprising an orally dissolvable and/or degradable composition and,optionally, one or more of the additional ingredients described herein.Alternately, formulations for buccal administration may comprise apowder and/or an aerosolized and/or atomized solution and/or suspensioncomprising the active ingredient. Such powdered, aerosolized, and/oraerosolized formulations, when dispersed, may have an average particleand/or droplet size in the range from about 0.1 to about 200 nanometers,and may further comprise one or more of the additional ingredientsdescribed herein.

A pharmaceutical composition described herein can be prepared, packaged,and/or sold in a formulation for ophthalmic administration. Suchformulations may, for example, be in the form of eye drops including,for example, a 0.1-1.0% (w/w) solution and/or suspension of the activeingredient in an aqueous or oily liquid carrier or excipient. Such dropsmay further comprise buffering agents, salts, and/or one or more otherof the additional ingredients described herein. Otherophthalmically-administrable formulations which are useful include thosewhich comprise the active ingredient in microcrystalline form and/or ina liposomal preparation. Ear drops and/or eye drops are alsocontemplated as being within the scope of this disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

Compounds provided herein are typically formulated in dosage unit formfor ease of administration and uniformity of dosage. It will beunderstood, however, that the total daily usage of the compositionsdescribed herein will be decided by a physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular subject or organism will depend upon a varietyof factors including the disease being treated and the severity of thedisorder; the activity of the specific active ingredient employed; thespecific composition employed; the age, body weight, general health,sex, and diet of the subject; the time of administration, route ofadministration, and rate of excretion of the specific active ingredientemployed; the duration of the treatment; drugs used in combination orcoincidental with the specific active ingredient employed; and likefactors well known in the medical arts.

The compounds and compositions provided herein can be administered byany route, including enteral (e.g., oral), parenteral, intravenous,intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by powders, ointments,creams, and/or drops), mucosal, nasal, bucal, sublingual; byintratracheal instillation, bronchial instillation, and/or inhalation;and/or as an oral spray, nasal spray, and/or aerosol. Specificallycontemplated routes are oral administration, intravenous administration(e.g., systemic intravenous injection), regional administration viablood and/or lymph supply, and/or direct administration to an affectedsite. In general, the most appropriate route of administration willdepend upon a variety of factors including the nature of the agent(e.g., its stability in the environment of the gastrointestinal tract),and/or the condition of the subject (e.g., whether the subject is ableto tolerate oral administration). In certain embodiments, the compoundor pharmaceutical composition described herein is suitable for topicaladministration to the eye of a subject.

The exact amount of a compound required to achieve an effective amountwill vary from subject to subject, depending, for example, on species,age, and general condition of a subject, severity of the side effects ordisorder, identity of the particular compound, mode of administration,and the like. An effective amount may be included in a single dose(e.g., single oral dose) or multiple doses (e.g., multiple oral doses).In certain embodiments, when multiple doses are administered to asubject or applied to a biological sample or cell, any two doses of themultiple doses include different or substantially the same amounts of acompound described herein. In certain embodiments, when multiple dosesare administered to a subject or applied to a biological sample or cell,the frequency of administering the multiple doses to the subject orapplying the multiple doses to the biological sample or cell is threedoses a day, two doses a day, one dose a day, one dose every other day,one dose every third day, one dose every week, one dose every two weeks,one dose every three weeks, or one dose every four weeks. In certainembodiments, the frequency of administering the multiple doses to thesubject or applying the multiple doses to the biological sample or cellis one dose per day. In certain embodiments, the frequency ofadministering the multiple doses to the subject or applying the multipledoses to the biological sample or cell is two doses per day. In certainembodiments, the frequency of administering the multiple doses to thesubject or applying the multiple doses to the biological sample or cellis three doses per day. In certain embodiments, when multiple doses areadministered to a subject or applied to a biological sample or cell, theduration between the first dose and last dose of the multiple doses isone day, two days, four days, one week, two weeks, three weeks, onemonth, two months, three months, four months, six months, nine months,one year, two years, three years, four years, five years, seven years,ten years, fifteen years, twenty years, or the lifetime of the subjector cell. In certain embodiments, the duration between the first dose andlast dose of the multiple doses is three months, six months, or oneyear. In certain embodiments, the duration between the first dose andlast dose of the multiple doses is the lifetime of the subject or cell.In certain embodiments, a dose (e.g., a single dose, or any dose ofmultiple doses) described herein includes independently between 0.1 μgand 1 μg, between 0.001 mg and 0.01 mg, between 0.01 mg and 0.1 mg,between 0.1 mg and 1 mg, between 1 mg and 3 mg, between 3 mg and 10 mg,between 10 mg and 30 mg, between 30 mg and 100 mg, between 100 mg and300 mg, between 300 mg and 1,000 mg, or between 1 g and 10 g, inclusive,of a compound described herein. In certain embodiments, a dose describedherein includes independently between 1 mg and 3 mg, inclusive, of acompound described herein. In certain embodiments, a dose describedherein includes independently between 3 mg and 10 mg, inclusive, of acompound described herein. In certain embodiments, a dose describedherein includes independently between 10 mg and 30 mg, inclusive, of acompound described herein. In certain embodiments, a dose describedherein includes independently between 30 mg and 100 mg, inclusive, of acompound described herein.

Dose ranges as described herein provide guidance for the administrationof provided pharmaceutical compositions to an adult. The amount to beadministered to, for example, a child or an adolescent can be determinedby a medical practitioner or person skilled in the art and can be loweror the same as that administered to an adult.

A compound or composition, as described herein, can be administered incombination with one or more additional pharmaceutical agents (e.g.,therapeutically and/or prophylactically active agents). The compounds orcompositions can be administered in combination with additionalpharmaceutical agents that improve their activity (e.g., activity (e.g.,potency and/or efficacy) in treating a disease in a subject in needthereof, in preventing a disease in a subject in need thereof, ininhibiting the activity of a kinase (e.g., BTK, HCK, LYN, BLK, FRK) in asubject, biological sample, or cell), improve bioavailability, improvesafety, reduce drug resistance, reduce and/or modify metabolism, inhibitexcretion, and/or modify distribution in a subject, biological sample,or cell. It will also be appreciated that the therapy employed mayachieve a desired effect for the same disorder, and/or it may achievedifferent effects. In certain embodiments, a pharmaceutical compositiondescribed herein including a compound described herein and an additionalpharmaceutical agent shows a synergistic effect that is absent in apharmaceutical composition including one of the compound and theadditional pharmaceutical agent, but not both.

The compound or composition can be administered concurrently with, priorto, or subsequent to one or more additional pharmaceutical agents, whichmay be useful as, e.g., combination therapies. Pharmaceutical agentsinclude therapeutically active agents. Pharmaceutical agents alsoinclude prophylactically active agents. Pharmaceutical agents includesmall organic molecules such as drug compounds (e.g., compounds approvedfor human or veterinary use by the U.S. Food and Drug Administration asprovided in the Code of Federal Regulations (CFR)), peptides, proteins,carbohydrates, monosaccharides, oligosaccharides, polysaccharides,nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides orproteins, small molecules linked to proteins, glycoproteins, steroids,nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides,antisense oligonucleotides, lipids, hormones, vitamins, and cells. Incertain embodiments, the additional pharmaceutical agent is apharmaceutical agent useful for treating and/or preventing a disease(e.g., proliferative disease, cancer, inflammatory disease, autoimmunedisease, genetic disease, hematological disease, neurological disease,painful condition, psychiatric disorder, or metabolic disorder) orpremalignant condition. Each additional pharmaceutical agent may beadministered at a dose and/or on a time schedule determined for thatpharmaceutical agent. The additional pharmaceutical agents may also beadministered together with each other and/or with the compound orcomposition described herein in a single dose or administered separatelyin different doses. The particular combination to employ in a regimenwill take into account compatibility of the compound described hereinwith the additional pharmaceutical agent(s) and/or the desiredtherapeutic and/or prophylactic effect to be achieved. In general, it isexpected that the additional pharmaceutical agent(s) in combination beutilized at levels that do not exceed the levels at which they areutilized individually. In some embodiments, the levels utilized incombination will be lower than those utilized individually.

The additional pharmaceutical agents include, but are not limited to,cytotoxic chemotherapeutic agents, epigenetic modifiers,glucocorticoids, immunotherapeutic agents, anti-proliferative agents,anti-cancer agents, anti-angiogenesis agents, anti-inflammatory agents,immunosuppressants, anti-bacterial agents, anti-viral agents,cardiovascular agents, cholesterol-lowering agents, anti-diabeticagents, anti-allergic agents, contraceptive agents, pain-relievingagents, and a combination thereof. In certain embodiments, theadditional pharmaceutical agent is an anti-proliferative agent (e.g.,anti-cancer agent). In certain embodiments, the additionalpharmaceutical agent is ABITREXATE (methotrexate), ADE, Adriamycin RDF(doxorubicin hydrochloride), Ambochlorin (chlorambucil), ARRANON(nelarabine), ARZERRA (ofatumumab), BOSULIF (bosutinib), BUSULFEX(busulfan), CAMPATH (alemtuzumab), CERUBIDINE (daunorubicinhydrochloride), CLAFEN (cyclophosphamide), CLOFAREX (clofarabine),CLOLAR (clofarabine), CVP, CYTOSAR-U (cytarabine), CYTOXAN(cyclophosphamide), ERWINAZE (Asparaginase Erwinia chrysanthemi),FLUDARA (fludarabine phosphate), FOLEX (methotrexate), FOLEX PFS(methotrexate), GAZYVA (obinutuzumab), GLEEVEC (imatinib mesylate),Hyper-CVAD, ICLUSIG (ponatinib hydrochloride), IMBRUVICA (ibrutinib),LEUKERAN (chlorambucil), LINFOLIZIN (chlorambucil), MARQIBO (vincristinesulfate liposome), METHOTREXATE LPF (methorexate), MEXATE(methotrexate), MEXATE-AQ (methotrexate), mitoxantrone hydrochloride,MUSTARGEN (mechlorethamine hydrochloride), MYLERAN (busulfan), NEOSAR(cyclophosphamide), ONCASPAR (Pegaspargase), PURINETHOL(mercaptopurine), PURIXAN (mercaptopurine), Rubidomycin (daunorubicinhydrochloride), SPRYCEL (dasatinib), SYNRIBO (omacetaxinemepesuccinate), TARABINE PFS (cytarabine), TASIGNA (nilotinib), TREANDA(bendamustine hydrochloride), TRISENOX (arsenic trioxide), VINCASAR PFS(vincristine sulfate), ZYDELIG (idelalisib), or a combination thereof.In certain embodiments, the additional pharmaceutical agent is ananti-lymphoma agent. In certain embodiments, the additionalpharmaceutical agent is ABITREXATE (methotrexate), ABVD, ABVE, ABVE-PC,ADCETRIS (brentuximab vedotin), ADRIAMYCIN PFS (doxorubicinhydrochloride), ADRIAMYCIN RDF (doxorubicin hydrochloride), AMBOCHLORIN(chlorambucil), AMBOCLORIN (chlorambucil), ARRANON (nelarabine),BEACOPP, BECENUM (carmazine), BELEODAQ (belinostat), BEXXAR (tositumomaband iodine 1131 tositumomab), BICNU (carmustine), BLENOXANE (bleomycin),CARMUBRIS (carmustine), CHOP, CLAFEN (cyclophosphamide), COPP, COPP-ABV,CVP, CYTOXAN (cyclophosphamide), DEPOCYT (liposomal cytarabine),DTIC-DOME (dacarbazine), EPOCH, FOLEX (methotrexate), FOLEX PFS(methotrexate), FOLOTYN (pralatrexate), HYPER-CVAD, ICE, IMBRUVICA(ibrutinib), INTRON A (recombinant interferon alfa-2b), ISTODAX(romidepsin), LEUKERAN (chlorambucil), LINFOLIZIN (chlorambucil),Lomustine, MATULANE (procarbazine hydrochloride), METHOTREXATE LPF(methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate), MOPP,MOZOBIL (plerixafor), MUSTARGEN (mechlorethamine hydrochloride), NEOSAR(cyclophosphamide), OEPA, ONTAK (denileukin diftitox), OPPA, R-CHOP,REVLIMID (lenalidomide), RITUXAN (rituximab), STANFORD V, TREANDA(bendamustine hydrochloride), VAMP, VELBAN (vinblastine sulfate),VELCADE (bortezomib), VELSAR (vinblastine sulfate), VINCASAR PFS(vincristine sulfate), ZEVALIN (ibritumomab tiuxetan), ZOLINZA(vorinostat), ZYDELIG (idelalisib), or a combination thereof. In certainembodiments, the additional pharmaceutical agent is REVLIMID(lenalidomide), DACOGEN (decitabine), VIDAZA (azacitidine), CYTOSAR-U(cytarabine), IDAMYCIN (idarubicin), CERUBIDINE (daunorubicin), LEUKERAN(chlorambucil), NEOSAR (cyclophosphamide), FLUDARA (fludarabine),LEUSTATIN (cladribine), ABITREXATE (methotrexate), ABRAXANE (paclitaxelalbumin-stabilized nanoparticle formulation), AC, AC-T, ADE, ADRIAMYCINPFS (doxorubicin hydrochloride), ADRUCIL (fluorouracil), AFINITOR(everolimus), AFINITOR DISPERZ (everolimus), ALDARA (imiquimod), ALIMTA(pemetrexed disodium), AREDIA (pamidronate disodium), ARIMIDEX(anastrozole), AROMASIN (exemestane), AVASTIN (bevacizumab), BECENUM(carmustine), BEP, BICNU (carmustine), BLENOXANE (bleomycin), CAF,CAMPTOSAR (irinotecan hydrochloride), CAPOX, CAPRELSA (vandetanib),CARBOPLATIN-TAXOL, CARMUBRIS (carmustine), CASODEX (bicalutamide), CEENU(lomustine), CERUBIDINE (daunorubicin hydrochloride), CERVARIX(recombinant HPV bivalent vaccine), CLAFEN (cyclophosphamide), CMF,COMETRIQ (cabozantinib-s-malate), COSMEGEN (dactinomycin), CYFOS(ifosfamide), CYRAMZA (ramucirumab), CYTOSAR-U (cytarabine), CYTOXAN(cyclophosphamide), DACOGEN (decitabine), DEGARELIX, DOXIL (doxorubicinhydrochloride liposome), DOXORUBICIN HYDROCHLORIDE, DOX-SL (doxorubicinhydrochloride liposome), DTIC-DOME (dacarbazine), EFUDEX (fluorouracil),ELLENCE (epirubicin hydrochloride), ELOXATIN (oxaliplatin), ERBITUX(cetuximab), ERIVEDGE (vismodegib), ETOPOPHOS (etoposide phosphate),EVACET (doxorubicin hydrochloride liposome), FARESTON (toremifene),FASLODEX (fulvestrant), FEC, FEMARA (letrozole), FLUOROPLEX(fluorouracil), FOLEX (methotrexate), FOLEX PFS (methotrexate), FOLFIRI,FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, FU-LV,GARDASIL (recombinant human papillomavirus (HPV) quadrivalent vaccine),GEMCITABINE-CISPLATIN, GEMCITABINE-OXALIPLATIN, GEMZAR (gemcitabinehydrochloride), GILOTRIF (afatinib dimaleate), GLEEVEC (imatinibmesylate), GLIADEL (carmustine implant), GLIADEL WAFER (carmustineimplant), HERCEPTIN (trastuzumab), HYCAMTIN (topotecan hydrochloride),IFEX (ifosfamide), IFOSFAMIDUM (ifosfamide), INLYTA (axitinib), INTRON A(recombinant interferon alfa-2b), IRESSA (gefitinib), IXEMPRA(ixabepilone), JAKAFI (ruxolitinib phosphate), JEVTANA (cabazitaxel),KADCYLA (ado-trastuzumab emtansine), KEYTRUDA (pembrolizumab), KYPROLIS(carfilzomib), LIPODOX (doxorubicin hydrochloride liposome), LUPRON(leuprolide acetate), LUPRON DEPOT (leuprolide acetate), LUPRON DEPOT-3MONTH (leuprolide acetate), LUPRON DEPOT-4 MONTH (leuprolide acetate),LUPRON DEPOT-PED (leuprolide acetate), MEGACE (megestrol acetate),MEKINIST (trametinib), METHAZOLASTONE (temozolomide), METHOTREXATE LPF(methotrexate), MEXATE (methotrexate), MEXATE-AQ (methotrexate),MITOXANTRONE HYDROCHLORIDE, MITOZYTREX (mitomycin c), MOZOBIL(plerixafor), MUSTARGEN (mechlorethamine hydrochloride), MUTAMYCIN(mitomycin c), MYLOSAR (azacitidine), NAVELBINE (vinorelbine tartrate),NEOSAR (cyclophosphamide), NEXAVAR (sorafenib tosylate), NOLVADEX(tamoxifen citrate), NOVALDEX (tamoxifen citrate), OFF, PAD, PARAPLAT(carboplatin), PARAPLATIN (carboplatin), PEG-INTRON (peginterferonalfa-2b), PEMETREXED DISODIUM, PERJETA (pertuzumab), PLATINOL(cisplatin), PLATINOL-AQ (cisplatin), POMALYST (pomalidomide),prednisone, PROLEUKIN (aldesleukin), PROLIA (denosumab), PROVENGE(sipuleucel-t), REVLIMID (lenalidomide), RUBIDOMYCIN (daunorubicinhydrochloride), SPRYCEL (dasatinib), STIVARGA (regorafenib), SUTENT(sunitinib malate), SYLATRON (peginterferon alfa-2b), SYLVANT(siltuximab), SYNOVIR (thalidomide), TAC, TAFINLAR (dabrafenib),TARABINE PFS (cytarabine), TARCEVA (erlotinib hydrochloride), TASIGNA(nilotinib), TAXOL (paclitaxel), TAXOTERE (docetaxel), TEMODAR(temozolomide), THALOMID (thalidomide), TOPOSAR (etoposide), TORISEL(temsirolimus), TPF, TRISENOX (arsenic trioxide), TYKERB (lapatinibditosylate), VECTIBIX (panitumumab), VEIP, VELBAN (vinblastine sulfate),VELCADE (bortezomib), VELSAR (vinblastine sulfate), VEPESID (etoposide),VIADUR (leuprolide acetate), VIDAZA (azacitidine), VINCASAR PFS(vincristine sulfate), VOTRIENT (pazopanib hydrochloride), WELLCOVORIN(leucovorin calcium), XALKORI (crizotinib), XELODA (capecitabine),XELOX, XGEVA (denosumab), XOFIGO (radium 223 dichloride), XTANDI(enzalutamide), YERVOY (ipilimumab), ZALTRAP (ziv-aflibercept), ZELBORAF(vemurafenib), ZOLADEX (goserelin acetate), ZOMETA (zoledronic acid),ZYKADIA (ceritinib), ZYTIGA (abiraterone acetate), ENMD-2076, PCI-32765,AC220, dovitinib lactate (TK1258, CHIR-258), BIBW 2992 (TOVOK™), SGX523,PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154,CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, and/orXL228), proteasome inhibitors (e.g., bortezomib (Velcade)), mTORinhibitors (e.g., rapamycin, temsirolimus (CCI-779), everolimus(RAD-001), ridaforolimus, AP23573 (Ariad), AZD8055, BEZ235, BGT226,XL765, PF-4691502, GDC0980, SF1126, and OSI-027), oblimersen,gemcitabine, carminomycin, leucovorin, pemetrexed, cyclophosphamide,dacarbazine, procarbizine, prednisolone, dexamethasone, campathecin,plicamycin, asparaginase, aminopterin, methopterin, porfiromycin,melphalan, leurosidine, leurosine, chlorambucil, trabectedin,procarbazine, discodermolide, carminomycin, aminopterin, and hexamethylmelamine, or a combination thereof. In certain embodiments, theadditional pharmaceutical agent is a cytotoxic chemotherapeutic agent(e.g., gemcitabine, cytarabine, daunorubicin, doxorubicin, vincristine,1-asparaginase, cyclophosphamide, or etoposide). In certain embodiments,the additional pharmaceutical agent is an epigenetic modifier such asazacitidine or romidepsin. In certain embodiments, the additionalpharmaceutical agent is ruxolitinib, BBT594, CHZ868, CYT387, orBMS911543. In certain embodiments, the additional pharmaceutical agentis an inhibitor of a tyrosine kinase. In some embodiments, theadditional pharmaceutical agent is a topoisomerase inhibitor, a MCL1inhibitor, a BCL-2 inhibitor, a BCL-xL inhibitor, a BRD4 inhibitor, aBRCA1 inhibitor, BRCA2 inhibitor, HER1 inhibitor, HER2 inhibitor, a CDK9inhibitor, a Jumonji histone demethylase inhibitor, or a DNA damageinducer. In certain embodiments, the additional pharmaceutical agent isa BCL-2 inhibitor. In certain embodiments, the additional pharmaceuticalagent is venetoclax. In some embodiments, the additional pharmaceuticalagent is etoposide, obatoclax, navitoclax, JQ1,4-(((5′-chloro-2′-(((1R,4R)-4-(((R)-1-methoxypropan-2-yl)amino)cyclohexyl)amino)-[2,4′-bipyridin]-6-yl)amino)methyl)tetrahydro-2H-pyran-4-carbonitrile,JIB04, or cisplatin. In certain embodiments, the additionalpharmaceutical agent is a binder or inhibitor of a kinase (e.g., JAK,ABL1, CDC2L1, CDC2L2, CSF1R, DDR1, DDR2, FLT4, KIT, PDGFRB, RET, TAOK2,or a combination thereof). In certain embodiments, the additionalpharmaceutical agent is an antibody or a fragment thereof (e.g.,monoclonal antibody). In certain embodiments, the additionalpharmaceutical agent is a tyrosine kinase inhibitor. In certainembodiments, the additional pharmaceutical agent is selected from thegroup consisting of epigenetic or transcriptional modulators (e.g., DNAmethyltransferase inhibitors, histone deacetylase inhibitors (HDACinhibitors), lysine methyltransferase inhibitors), antimitotic drugs(e.g., taxanes and vinca alkaloids), hormone receptor modulators (e.g.,estrogen receptor modulators and androgen receptor modulators), cellsignaling pathway inhibitors (e.g., tyrosine protein kinase inhibitors),modulators of protein stability (e.g., proteasome inhibitors), Hsp90inhibitors, glucocorticoids, all-trans retinoic acids, and other agentsthat promote differentiation. In certain embodiments, the additionalpharmaceutical agent is a glucocorticoid (e.g., cortisol, cortisone,prednisone, methylprednisolone, dexamethasone, betamethasone,triamcinolone, fludrocortisone acetate, or deoxycorticosterone acetate).In certain embodiments, the additional therapy is an immunotherapy(e.g., an immunotherapeutic monoclonal antibody). In certainembodiments, the additional pharmaceutical agent is an immunomodulator.In certain embodiments, the additional pharmaceutical agent is an immunecheckpoint inhibitor. In certain embodiments, the additionalpharmaceutical agent is a programmed cell death 1 protein (PD-1)inhibitor. In certain embodiments, the additional pharmaceutical agentis a programmed cell death 1 protein ligand 1 (PD-L1) inhibitor. Incertain embodiments, the additional pharmaceutical agent is a cytotoxicT-lymphocyte-associated protein 4 (CTLA-4) inhibitor. In certainembodiments, the additional pharmaceutical agent is a BTK inhibitor(e.g., ibrutinib, CC-292, ONO-4059, evobrutinib, spebrutinib, BGB-3111,HM71224, or ACP-196 (i.e., acalabrutinib)). In certain embodiments, thecompounds described herein or pharmaceutical compositions can beadministered in combination with an anti-cancer therapy including, butnot limited to, surgery, radiation therapy, and transplantation (e.g.,stem cell transplantation, bone marrow transplantation).

In certain embodiments, the additional pharmaceutical agent is a BCL-2inhibitor (e.g., venetoclax, navitoclax, obatoclax), a BCL-2/BCL-xLinhibitor (e.g., APG-1252, BM-1197).

In certain embodiments, the additional pharmaceutical agent isvenetoclax.

Also encompassed by the present disclosure are kits (e.g.,pharmaceutical packs). In certain embodiments, the kit comprises acompound or pharmaceutical composition described herein, andinstructions for using the compound or pharmaceutical composition. Incertain embodiments, the kit comprises a first container, wherein thefirst container includes the compound or pharmaceutical composition. Insome embodiments, the kit further comprises a second container. Incertain embodiments, the second container includes an excipient (e.g.,an excipient for dilution or suspension of the compound orpharmaceutical composition). In certain embodiments, the secondcontainer includes an additional pharmaceutical agent. In someembodiments, the kit further comprises a third container. In certainembodiments, the third container includes an additional pharmaceuticalagent. In some embodiments, the compound or pharmaceutical compositionincluded in the first container and the excipient or additionalpharmaceutical agent included in the second container are combined toform one unit dosage form. In some embodiments, the compound orpharmaceutical composition included in the first container, theexcipient included in the second container, and the additionalpharmaceutical agent included in the third container are combined toform one unit dosage form. In certain embodiments, each of the first,second, and third containers is independently a vial, ampule, bottle,syringe, dispenser package, tube, or inhaler.

In certain embodiments, the instructions are for administering thecompound or pharmaceutical composition to a subject (e.g., a subject inneed of treatment or prevention of a disease described herein). Incertain embodiments, the instructions are for contacting a biologicalsample or cell with the compound or pharmaceutical composition. Incertain embodiments, the instructions comprise information required by aregulatory agency, such as the U.S. Food and Drug Administration (FDA)or the European Agency for the Evaluation of Medicinal Products (EMA).In certain embodiments, the instructions comprise prescribinginformation.

The compounds, pharmaceutical compositions, and kits described hereinmay synergistically augment inhibition of a kinase (e.g., BTK, HCK, LYN,BLK, FRK) induced by the additional pharmaceutical agent(s) in thebiological sample or subject. Thus, the combination of the compounds,pharmaceutical compositions, or kits with additional pharmaceuticalagent(s) may be useful in treating diseases resistant to a treatmentusing the additional pharmaceutical agent(s) without the compounds,pharmaceutical compositions, or kits described herein.

Methods of Treatment and Uses

The present disclosure provides methods of modulating (e.g., inhibitingor increasing) the activity (e.g., aberrant activity, such as increasedor decreased activity) of a kinase (e.g., BTK, HCK, LYN). The presentdisclosure also provides methods of modulating (e.g., inhibiting orincreasing) the activity (e.g., undesired or aberrant activity, such asincreased activity (e.g., activity above normal levels) or decreasedactivity (e.g., activity below normal levels)), of a kinase in asubject, biological sample, or cell. The present disclosure alsoprovides methods for the treatment of a range of diseases andconditions, such as diseases and conditions associated with undesired oraberrant activity (e.g., increased activity) or overexpression of akinase (e.g., BTK, HCK, LYN). In certain embodiments, the diseasesinclude a proliferative disease (e.g., an IgM gammopathy (e.g., an IgMMonoclonal gammopathy of undetermined significance (MGUS), amyloid lightchain (AL) amyloidosis), mastocytosis (e.g., systemic mastocytosis),cancer (e.g., breast cancer, colon cancer, testicular cancer, CNScancer, stomach cancer, lymphoma (e.g., B-cell lymphoma (e.g.,lymphoplasmacytic lymphoma (e.g., IgM secreting lymphoplasmacyticlymphoma (i.e., Waldenström's Macroglobulinemia), non-IgM secretinglymphoplasmacytic lymphoma)), diffuse large B-cell lymphoma (e.g.,activated B-cell-like (ABC)-DLBCL, germinal center B-cell-like(GBC)-DLBCL), follicular lymphoma, marginal zone B-cell lymphoma, smalllymphocytic lymphoma, mantle cell lymphoma), myeloma (e.g., IgM myelomas(e.g., IgM multiple myeloma)), and leukemia (e.g., chronic lymphocyticleukemia (CLL), acute lymphoblastic leukemia, myelogenous leukemia(e.g., chronic myelogenous leukemia, acute myelogenous leukemia (e.g.,mast cell leukemia), myeloproliferative diseases (e.g., myelodysplasticsyndrome))))).

In another aspect, the present disclosure provides methods of treating adisease in a subject in need thereof, the method comprisingadministering to the subject in need thereof an effective amount (e.g.,therapeutically effective amount) of a compound described herein or apharmaceutical composition described herein.

In another aspect, the present disclosure provides methods of preventinga disease in a subject in need thereof, the method comprisingadministering to the subject in need thereof an effective amount (e.g.,prophylactically effective amount) of a compound described herein or apharmaceutical composition described herein.

In another aspect, the present disclosure provides methods of inhibitingthe activity of a kinase in a subject in need thereof, the methodcomprising administering to the subject in need thereof an effectiveamount of a compound described herein or a pharmaceutical compositiondescribed herein.

In another aspect, the present disclosure provides methods of inhibitingthe activity of a kinase in a biological sample (e.g., an in vitrobiological sample), the method comprising contacting the biologicalsample with an effective amount of a compound described herein or apharmaceutical composition described herein.

In another aspect, the present disclosure provides methods of inhibitingthe activity of a kinase in a cell (e.g., an in vitro cell), the methodcomprising contacting the cell with an effective amount of a compounddescribed herein or a pharmaceutical composition described herein.

Without wishing to be bound by any particular theory, in certainembodiments the compounds described herein are able to bind (e.g.,covalently modify) the kinase being inhibited. In certain embodiments, acompound described herein is able to bind (e.g., covalently modify) tothe kinase. In certain embodiments, the kinase is HCK. In certainembodiments, the kinase is BTK. In certain embodiments, the kinase isLYN.

In certain embodiments, provided are methods of decreasing the activityof a kinase (e.g., HCK, BTK, LYN) in a subject, biological sample, orcell by at least about 1%, at least about 3%, at least about 10%, atleast about 20%, at least about 30%, at least about 40%, at least about50%, at least about 60%, at least about 70%, at least about 80%, or atleast about 90%. In certain embodiments, the activity of a kinase in asubject, biological sample, or cell is decreased by at least about 1%,at least about 3%, at least about 10%, at least about 20%, at leastabout 30%, at least about 40%, at least about 50%, at least about 60%,at least about 70%, at least about 80%, or at least about 90%. In someembodiments, the activity of a kinase in a subject, biological sample,or cell is selectively inhibited by the method. In some embodiments, theactivity of a kinase (e.g., HCK, BTK, LYN) in a subject, biologicalsample, or cell is selectively decreased by a compound or pharmaceuticalcomposition described herein.

A disease, including proliferative disease, may be associated withaberrant or undesired activity of a kinase, and/or overexpression of thekinase. Aberrant or undesired activity of a kinase may be an increasedor a decreased level of activity of the kinase. Proliferative diseasesare sometimes associated with abnormal levels of HCK, BTK, or LYNactivity, frequently through increased or decreased HCK, BTK, or LYNactivation. Inhibition of the activity of HCK, BTK, or LYN would beexpected to inhibit phosphorylation. In certain embodiments, HCK, BTK,or LYN is not overexpressed, but the activity of HCK, BTK, or LYN isincreased. In certain embodiments, HCK, BTK, or LYN is overexpressed,and the activity of HCK, BTK, or LYN is increased. The compounds andpharmaceutical compositions described herein may inhibit the activity ofHCK, BTK, or LYN and be useful in treating and/or preventing diseases,such as diseases associated with the aberrant, increased, or undesiredactivity of a kinase, over activation of the kinase, and/oroverexpression of the kinase.

In certain embodiments, the disease (e.g., the disease to be treated orprevented by a method described herein) is associated with the increasedactivity of a kinase (e.g., HCK, BTK, LYN). In certain embodiments, thedisease is associated with overexpression of a kinase (e.g., HCK, BTK,LYN). In certain embodiments, the disease is a proliferative disease. Incertain embodiments, the proliferative disease is cancer. In certainembodiments, the cancer is associated with a mutation in MYD88. Inanother embodiment, the cancer is associated with mutated BTK. Incertain embodiments, the proliferative disease is mastocytosis. Incertain embodiments, the mastocytosis is systemic mastocytosis. Incertain embodiments, the proliferative disease is an IgM gammopathy. Incertain embodiments, the IgM gammopathy is IgM monoclonal gammopathywith undetermined significance.

In certain embodiments, the cancer is breast cancer. In certainembodiments, the cancer is colon cancer. In certain embodiments, thecancer is testicular cancer. In certain embodiments, the cancer is CNScancer. In certain embodiments, the cancer is stomach cancer. In certainembodiments, the cancer is lymphoma. In certain embodiments, thelymphoma is a B-cell Lymphoma. In certain embodiments, the B-celllymphoma is lymphoplasmacytic lymphoma. In certain embodiments, thelymphoplasmacytic lymphoma is IgM secreting lymphoplasmacytic lymphoma(i.e., Waldenström's macroglobulinemia). In certain embodiments, thelymphoplasmacytic lymphoma is non-IgM secreting. In certain embodiments,the B-cell lymphoma is Diffuse Large B-Cell Lymphoma (DLBCL). In certainembodiments, the DLBCL is activated B-cell-like (ABC)-DLBCL. In certainembodiments, the DLBCL is germinal center B-cell-like (GBC)-DLBCL. Incertain embodiments, the lymphoma is follicular lymphoma. In certainembodiments, the lymphoma is marginal zone B-cell lymphoma. In certainembodiments, the lymphoma is small lymphocytic lymphoma. In certainembodiments, the small lymphocytic lymphoma is mantle cell lymphoma. Incertain embodiments, the cancer is leukemia. In certain embodiments, theleukemia is chronic lymphocytic leukemia (CLL). In certain embodiments,the leukemia is myelogenous leukemia. In certain embodiments, themyelogenous leukemia is chronic myelogenous leukemia. In certainembodiments, the myelogenous leukemia is acute myelogenous leukemia. Incertain embodiments, the acute myelogenous leukemia is mast cellleukemia. In certain embodiments, the cancer is myeloma. In certainembodiments. the myeloma is IgM myeloma. In certain embodiments, the IgMmyeloma is IgM multiple myeloma. In certain embodiments, the cancer is amyeloproliferative disease. In certain embodiments, themyeloproliferative disease is myelodysplastic syndrome.

Further provided herein is a method of treating a subject comprisingadministering to a subject with an MYD88 mutated disease. An MYD88mutated disease can include, but is not limited to a proliferativedisease (e.g., an IgM gammopathy (e.g., an IgM Monoclonal gammopathy ofundetermined significance (MGUS), amyloid light chain (AL) amyloidosis),mastocytosis (e.g., systemic mastocytosis), cancer (e.g., breast cancer,colon cancer, testicular cancer, CNS cancer, stomach cancer, lymphoma(e.g., B-cell lymphoma (e.g., lymphoplasmacytic lymphoma (e.g., IgMsecreting lymphoplasmacytic lymphoma (i.e., Waldenström'sMacroglobulinemia), non-IgM secreting lymphoplasmacytic lymphoma)),diffuse large B-cell lymphoma (e.g., activated B-cell-like (ABC)-DLBCL,germinal center B-cell-like (GBC)-DLBCL), follicular lymphoma, marginalzone B-cell lymphoma, small lymphocytic lymphoma, mantle cell lymphoma),myeloma (e.g., IgM myelomas (e.g., IgM multiple myeloma)), and leukemia(e.g., chronic lymphocytic leukemia (CLL), acute lymphoblastic leukemia,myelogenous leukemia (e.g., chronic myelogenous leukemia, acutemyelogenous leukemia (e.g., mast cell leukemia), myeloproliferativediseases (e.g., myelodysplastic syndrome))))).

In certain embodiments, the method described herein is superior (e.g.,showing improved safety and/or therapeutic effects; decreased adverseeffects) or comparable to existing therapy (e.g., chemotherapy).

In certain embodiments, the biological sample or cell (e.g., thebiological sample or cell being contacted with a compound orpharmaceutical composition described herein) is in vitro. In certainembodiments, the cell is in vivo. In certain embodiments, the biologicalsample or cell is ex vivo.

In certain embodiments, the cell is a malignant cell (e.g., cancercell). In certain embodiments, the cell is a malignant blood cell. Incertain embodiments, the cell is a malignant bone marrow cell. Incertain embodiments, the cell is a malignant white blood cell.

In certain embodiments, the cell is an adenocarcinoma cell, blastomacell, carcinoma cell, or sarcoma cell. In certain embodiments, the cellis a pre-malignant cell (e.g., pre-cancerous cell).

In certain embodiments, the method described herein further comprisesadministering to the subject in need thereof an additional therapy. Incertain embodiments, the additional therapy is an additionalpharmaceutical agent described herein. In certain embodiments, theadditional therapy is a cytotoxic chemotherapy (e.g., gemcitabine,cytarabine, daunorubicin, doxorubicin, vincristine, 1-asparaginase,cyclophosphamide, or etoposide). In certain embodiments, the additionaltherapy is an epigenetic modifier (e.g., azacitidine or romidepsin). Incertain embodiments, the additional therapy is a glucocorticoid.

In certain embodiments, the additional therapy is an immunotherapy(e.g., an immunotherapeutic monoclonal antibody). In some embodiments,the additional pharmaceutical agent is etoposide, obatoclax, ornavitoclax, and optionally the disease is breast cancer, e.g.,triple-negative breast cancer, HER2 positive breast cancer, HER2negative breast cancer, ER-positive breast cancer, ER-negative breastcancer, or ER/PR-positive breast cancer. In some embodiments, theadditional pharmaceutical agent is etoposide, JIB04, or cisplatin. Insome embodiments, the additional pharmaceutical agent is JQ1 or NVP2,and optionally the disease is leukemia, e.g., acute myelogenousleukemia, myeloblastic leukemia, promyelocytic leukemia, myelomonocyticleukemia, monocytic leukemia, monoblastic leukemia, or megakaryoblasticleukemia.

In yet another aspect, the present disclosure provides compounds andpharmaceutical compositions described herein for use in the treatment ofa disease (e.g., a proliferative disease, such as an IgM gammopathy,mastocytosis, or cancer) in a subject in need thereof.

In yet another aspect, the present disclosure provides compounds andpharmaceutical compositions described herein for use in the preventionof a disease (e.g., a proliferative disease, such as an IgM gammopathy,mastocytosis, or cancer) in a subject in need thereof.

In another aspect, the present disclosure provides compounds andpharmaceutical compositions described herein for use in inhibiting theactivity of a kinase (e.g., SRC Family kinases (e.g., HCK, LYN, BLK,FRK), Tec family kinases (e.g., BTK)) in a subject in need thereof.

In another aspect, the present disclosure provides compounds andpharmaceutical compositions described herein for use in inhibiting theactivity of a kinase in a biological sample (e.g., an in vivo or ex vivobiological sample).

In another aspect, the present disclosure provides compounds andpharmaceutical compositions described herein for use in inhibiting theactivity of a kinase in a cell (e.g., an in vivo or ex vivo cell).

In another aspect, the present disclosure provides uses of compounds andpharmaceutical compositions described herein in the manufacture of amedicament for treating a disease in a subject in need thereof.

In another aspect, the present disclosure provides uses of compounds andpharmaceutical compositions described herein in the manufacture of amedicament for preventing a disease in a subject in need thereof.

EXAMPLES

In order that the disclosure described herein may be more fullyunderstood, the following examples are set forth. The synthetic andbiological examples described in this application are offered toillustrate the compounds, pharmaceutical compositions, and methodsprovided herein and are not to be construed in any way as limiting theirscope.

Example 1. Dose Response Curves for HCK Inhibitors

In vitro cellular efficacies (drug dose-response) were measured byCellTiter-Glo™ cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WMcells; TMD8, HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19GCB DLBCL cells; Ramos Burkitt's lymphoma cells; and multiple myelomacells. Cells were treated with either Compound I-4 (FIG. 2 ) or CompoundI-1 (FIG. 3 ) with serially diluted drugs for 72 hrs (FIG. 2 ).

Example 2. Dose Response Curves for HCK Inhibitors

In vitro cellular efficacies (drug dose-response) of ibrutinib orCompound I-4 were measured by CellTiter-Glo™ cell viability assay invector only, BTK WT or BTK C481S expressing lentiviral vector transducedTMD8 ABC-DLBCL or BCWM.1 WM cells. Cells were treated with seriallydiluted ibrutinib or SB1-G-114 for 72 hrs. The results show thatCompound I-4 overcomes BTKC481S mediated ibrutinib resistance in MYD88mutated WM and ABC DLBCL cells (FIG. 4 ).

Example 3. ED₅₀ Values

ED₅₀ values of the different compounds were measured by CellTiter-Glo™cell viability assay in MYD88 mutated BCWM.1, MWCL-1 WM cells; TMD8,HBL-1 ABC DLBCL cells; MYD88 wild type OCI-Ly7, OCI-Ly19 GCB DLBCLcells; Ramos Burkitt's lymphoma cells; and RPMI-8226 multiple myelomacells. Cells were treated with serially diluted drugs for 72 hrs

TABLE 1 ED₅₀ values of compounds # BCWM.1 MWCL-1 TMD8 HBL-1 OCI-Ly7OCI-Ly19 Ramos RPMI-8226 cis I-1 7.50E−07 9.86E−07 8.26E−07 5.88E−062.31E−06 4.46E−06 4.44E−06 2.36E−05 trans I-1 9.99E−09 1.00E−08 3.52E−083.59E−07 4.33E−08 8.59E−06 1.15E−07 8.02E−05 cis I-3 6.51E−07 8.43E−076.24E−07 1.01E−06 7.94E−07 1.17E−06 8.10E−07 1.02E−06 trans I-3 9.73E−071.33E−06 6.84E−07 1.58E−06 7.11E−07 8.38E−07 8.81E−07 2.12E−06 cis I-41.00E−06 8.10E−07 7.57E−07 1.60E−06 1.36E−06 1.70E−06 1.31E−06 3.05E−06trans I-4 1.09E−07 2.28E−08 9.54E08  5.28E−07 8.28E−07 1.96E−06 8.94E−073.11E−06 I-5 4.39E−06 2.30E−06 2.99E−06 1.27E−05 7.48E−06 1.61E−052.84E−05 4.27E−05 trans I-6 4.24E−07 1.40E−07 7.19E−07 2.23E−05 7.54E−072.33E−05 8.56E−05 1.49E−02 cis I-6 2.47E−06 1.21E−06 1.75E−06 5.89E−065.24E−06 9.14E−06 7.32E−06 1.76E−05 trans I-7 N 8.85E−05 2.37E−071.83E−06 1.73E−06 1.16E−06 8.73E−06 1.40E−05 cis I-7 4.18E−07 2.95E−062.71E−07 6.13E−06 3.97E−06 4.85E−06 7.20E−06 3.40E−05 trans I-8 7.14E−064.91E−06 4.80E−06 6.81E−06 3.39E−06 6.24E−06 6.87E−05 2.14E−05 cis I-81.57E−05 1.41E−05 N 1.68E−05 2.09E−05 1.44E−05 4.33E−05 2.40E−05 transI-9 N N 3.32E−06 7.63E−06 2.79E−04 N N N cis I-9 5.38E−06 N 6.44E−067.98E−06 5.99E−06 5.37E−06 1.61E−05 1.20E−04 trans I-10 8.04E−073.50E−06 1.56E−05 8.61E−06 2.15E−06 N 4.21E−06 3.02E−04 cis I-101.06E−06 1.02E−06 8.06E−07 1.85E−06 1.10E−06 1.53E−06 6.58E−07 1.38E−06

Example 4. Synthesis of Cis and Trans I-4

cis1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine(cis I-4) and trans1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine(trans I-4) were synthesized according to the following scheme.

4-chloro-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridine(1)

A mixture of 4-chloro-3-iodo-1H-pyrazolo[4,3-c]pyridine (3 g, 10.7mmol), 1,4-dioxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate (5 g, 16.1mmol), and Cs₂CO₃ (7 g, 21.4 mmol) in DMF (100 mL) was stirred at 90° C.for 16 h. After the reaction was complete, the mixture was concentratedand purified by silica gel (DCM/MeOH=10/1) to obtain compound 1 (whitesolid, 700 mg, 16% yield). LCMS: 420 (M+H)⁺.

N-(2,4-dimethoxybenzyl)-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine(2)

A mixture of4-chloro-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridine1 (400 mg, 0.95 mmol), (2,4-dimethoxyphenyl)methanamine (319 mg, 1.91mmol), and Cs₂CO₃ (620 mg, 1.91 mmol) was stirred at 125° C. for 16 h.After the reaction was complete, the mixture was concentrated andpurified by silica gel (DCM/MeOH=10/1) to obtain compound 2 (yellowsolid, 500 mg, 95% yield). LCMS: 551 (M+H)⁺.

N-(2,4-dimethoxybenzyl)-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine(3)

A mixture ofN-(2,4-dimethoxybenzyl)-3-iodo-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine2 (250 mg, 0.45 mmol), K₂CO₃ (120 mg, 0.9 mmol), Pd(PPh₃)₂Cl₂ (150 mg,0.45 mmol), and4,4,5,5-tetramethyl-2-(4-phenoxyphenyl)-1,3,2-dioxaborolane (130 mg,0.45 mmol) in THF (5 mL) and H₂O (0.1 mL) was stirred at 60° C. for 16h. After the reaction was complete the mixture was concentrated andpurified by prep-HPLC (C18 column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃)to get title compound 3 (white solid, 170 mg, 63%). LCMS: 593 (M+H)⁺.

4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-1-yl)cyclohexanone(4)

The mixture ofN-(2,4-dimethoxybenzyl)-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[4,3-c]pyridin-4-amine3 (238 mg, 0.4 mmol) in TFA (5 mL), and DCM (2 mL) was stirred at 45° C.for 16 h. After the reaction was complete, the mixture was concentratedand purified by prep-HPLC (C18 column, CH₃CN/H₂O, containing 0.05%NH₄HCO₃) to get compound 4 (white solid, 150 mg, 94%). LCMS: 399 (M+H)⁺.

cis1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine(cis I-4) and trans1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-4-amine(trans I-4)

A mixture of4-(4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[4,3-c]pyridin-1-yl)cyclohexanone (150 mg, 0.25 mmol), 1-methylpiperazine (100 mg, 1.00mmol), and NaBH(OAc)₃ (212 mg, 1.00 mmol) in THF (5 mL) was stirred atrt for 16 h. After the reaction was complete the mixture wasconcentrated and purified by prep-HPLC (C18 column, CH₃CN/H₂O,containing 0.05% NH₄HCO₃) to get title compound cis I-4 (white solid, 4mg, 2.2%) and trans I-4 (white solid, 3 mg, 2.1%). HPLC: 100% (@254 nm);

cis I-4: LCMS: 483 (M+H)⁺; ¹H NMR (400 MHz, CDCl₃) δ 7.83 (d, J=6.2 Hz,1H), 7.75-7.55 (m, 2H), 7.37 (tt, J=39.2, 19.7 Hz, 2H), 7.21-6.84 (m,5H), 6.82 (d, J=6.2 Hz, 1H), 4.97 (s, 2H), 4.71-4.20 (m, 1H), 3.00-2.34(m, 9H), 2.33-2.12 (m, 7H), 1.82-1.68 (m, 2H), 1.60 (dd, J=14.0, 11.1Hz, 2H).

trans I-4: LCMS: 483 (M+H)⁺; ¹H NMR (400 MHz, CDCl₃) δ 7.83 (d, J=6.2Hz, 1H), 7.70-7.56 (m, 2H), 7.46-7.32 (m, 2H), 7.20-6.97 (m, 5H), 6.72(d, J=6.3 Hz, 1H), 5.17 (d, J=103.3 Hz, 2H), 4.58-4.07 (m, 1H), 2.67 (s,4H), 2.59-2.34 (m, 5H), 2.30 (s, 3H), 2.23-1.92 (m, 6H), 1.51 (dd,J=11.7, 4.5 Hz, 2H).

Example 5. Synthesis of Cis and Trans I-3

Synthesis of cis1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrrolo[3,2-c]pyridin-4-amine(cis I-3) and trans1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrrolo[3,2-c]pyridin-4-amine(trans I-3) were synthesized following the same procedure as cis andtrans I-4, but starting with 4-chloro-3-iodo-1H-pyrrolo[3,2-c]pyridineinstead.

cis I-3: LCMS (m/z): 482 [M+H]⁺; ¹H NMR (400 MHz, DMSO) δ 7.73 (d, J=6.4Hz, 2H), 7.46 (t, J=8.8 Hz, 2H), 7.38 (t, J=8.0 Hz, 2H), 7.15 (t, J=7.6Hz, 1H), 7.07-7.10 (m, 5H), 6.83 (d, J=6.0 Hz, 1H), 5.09 (br, 2H), 4.26(m, 1H), 2.48-2.51 (m, 7H), 2.31 (m, 4H), 2.13-2.25 (m, 5H), 1.55-1.62(m, 3H), 1.25 (m, 1H).

trans I-3: LCMS (m/z): 482 [M+H]+; ¹H NMR (400 MHz, DMSO) δ 7.72 (d,J=6.4 Hz, 2H), 7.43 (t, J=8.4 Hz, 2H), 7.38 (t, J=8.0 Hz, 2H), 7.15 (t,J=7.6 Hz, 1H), 7.01-7.08 (m, 5H), 6.75 (d, J=6.4 Hz, 1H), 5.27 (br, 2H),4.14 (m, 1H), 2.40-2.67 (m, 8H), 2.31 (s, 3H), 2.12-2.25 (m, 4H),1.53-1.59 (m, 3H), 1.31 (m, 2H).

Example 6. Synthesis of Cis and Trans I-5

6-amino-9-(4-(4-methylpiperazin-1-yl)cyclohexyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one(I-5) was synthesized according to the following scheme.

N,N-dibenzyl-6-chloro-5-nitropyrimidin-4-amine (1a)

To a mixture of 4,6-dichloro-5-nitropyrimidine (10 g, 51.8 mmol) andDIPEA (13.4 g, 103.6 mmol) in DCM (40 mL) was added dropwise a solutionof dibenzylamine (10.2 g, 51.8 mmol) in DCM (20 mL) at 0° C., and themixture was stirred at rt. After 16 h, the reaction was concentrated andpurified by column chromatography (silica gel, EA/PE=1/6) to afford thedesired product 1a as yellow solid (15.0 g, yield 90%). LCMS (m/z):355.0 [M+H]⁺.

N⁴,N⁴-dibenzyl-5-nitro-N⁶-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,6-diamine(2a)

To the mixture of N,N-dibenzyl-6-chloro-5-nitropyrimidin-4-amine 1 (15g, 42.3 mmol) and DIPEA (10.2 g, 84.6 mmol) in DCM (40 mL) was addeddropwise the solution of 1,4-dioxaspiro[4.5]decan-8-amine (6.6 g, 42.3mmol) in DCM (20 mL), the mixture was stirred at rt for 16 h,concentrated and purified by column chromatography (silica gel,EA/PE=1/2) to afford desired product 2a as yellow solid (13.0 g, yield65%). LCMS (m/z): 476.1 [M+H]+.

N⁴,N⁴-dibenzyl-N⁶-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,5,6-triamine(3a)

A mixture ofN⁴,N⁴-dibenzyl-5-nitro-N⁶-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,6-diamine2a (10.0 g, 21.0 mmol), Fe (5.8 g, 105.0 mmol) and NH₄Cl (5.9 g, 105.0mmol) in EtOH (50 mL) was stirred at 80° C. After 3 h, the mixture wasfiltered, and the filtrate was concentrated to leave crude product 3a asyellow solid (8.0 g crude, yield 86%). LCMS (m/z): 446.1 [M+H]⁺.

6-(dibenzylamino)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7,9-dihydro-8H-purin-8-one(4a)

To a mixture ofN⁴,N⁴-dibenzyl-N6-(1,4-dioxaspiro[4.5]decan-8-yl)pyrimidine-4,5,6-triamine3a (6.0 g, 13.4 mmol) and DIPEA (5.0 mL, 26.8 mmol) in DCM (60 mL) wasadded Triphosgene (bis(trichloromethyl) carbonate (BTC) (1.6 g, 5.4mmol) at 0° C. The mixture was then stirred at 40° C. for 3 h,concentrated, and purified by column chromatography (silica gel,MeOH/DCM=1/20) to afford product 4a as yellow solid (3.0 g, yield 47%).LCMS (m/z): 472.1 [M+H]⁺.

6-amino-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7,9-dihydro-8H-purin-8-one(5a)

A mixture of6-(dibenzylamino)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-purin-8(9H)-one4a (2.5 g, 5.3 mmol) and Pd/C (10%, 0.3 g) in EtOH (50 mL) was stirredat 60° C. under hydrogen (1 atm) for 16 h. The mixture was then filteredthrough celite, and the filtrate was concentrated and purified by columnchromatography (silica gel, EA/PE=3/1) to afford product 5a as yellowsolid (1.1 g, yield 73%). LCMS (m/z): 292.0 [M+H]⁺.

6-amino-7-(4-phenoxyphenyl)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7,9-dihydro-8H-purin-8-one(6a)

A mixture of6-amino-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-purin-8(9H)-one 5 (1.1 g,3.8 mmol), (4-phenoxyphenyl)boronic acid (1.6 g, 7.6 mmol), Cu(OAc)₂(1.4 g, 7.6 mmol) and DIPEA (1.3 mL, 7.6 mmol) in DCM (30 mL) wasstirred at 40° C. for 16 h. The mixture was then filtered, and thefiltrate was concentrated and purified by column chromatography (silicagel, MeOH/DCM=1/20) to obtain product 6a as yellow solid (0.8 g, yield47%). LCMS (m/z): 460.1 [M+H]⁺.

6-amino-9-(4-oxocyclohexyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one(7a)

A mixture of6-amino-7-(4-phenoxyphenyl)-9-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-purin-8(9H)-one6a (0.7 g, 1.5 mmol) and concentrated HCl solution (8.0 mL) in acetone(12 mL) was stirred at rt for 3 h. The mixture was then concentratedunder reduced pressure, and the residue was diluted with water (10 mL).The solution was adjusted to pH 8 with Na₂CO₃ and extracted with ethylacetate (150 mL×2), and the combined organic was dried over anhydrousNa₂SO₄, filtered and concentrated to leave crude product 7a as yellowsolid (0.8 g).

6-amino-9-(4-(4-methylpiperazin-1-yl)cyclohexyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one(I-5)

A mixture of6-amino-9-(4-oxocyclohexyl)-7-(4-phenoxyphenyl)-7H-purin-8(9H)-one 7(0.5 g, 1.2 mmol), 1-methylpiperazine (2.4 g, 24 mmol) and CH₃COOH (0.1mL) in DCE (10 mL) was stirred at rt for 3 h. After this time,NaBH(OAc)₃ (1.0 g, 4.8 mmol) was added, and the mixture was stirred atrt overnight and concentrated in vacuum. The residue was then dilutedwith ethyl acetate (150 mL), washed with brine (50 mL×2), dried overanhydrous Na₂SO₄, filtered, concentrated and purified by preparativeHPLC (C18 column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃) to afford I-5 aswhite solid (64.1 mg, yield 10%). LCMS (m/z): 500.1 [M+H]⁺; ¹H NMR(DMSO-d₆, 400 MHz): δ (ppm) 8.12 (s, 1H), 7.45-7.41 (m, 4H), 7.19 (t,J=7.6 Hz, 1H), 7.13 (t, J=6.4 Hz, 4H), 5.73 (s, 2H), 4.32 (t, J=12.0 Hz,1H), 2.67-2.57 (m, 3H), 2.35 (m, 6H), 2.15 (s, 3H), 2.12-2.06 (m, 4H),1.47-1.41 (m, 4H).

Example 7. Synthesis of Cis and Trans I-6

The synthesis of trans4-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(trans I-6) and cis4-amino−1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(cis I-6) was carried out according to the following scheme.

2-chloro-3-nitro-N-(1,4-dioxaspiro[4.5]decan-8-yl)pyridin-4-amine (1b

To a mixture of 2,4-dichloro-3-nitropyridine (8.5 g, 44.2 mmol) andDIPEA (11.4 g, 88.4 mmol) in EtOH (40 mL) was added dropwise a solutionof 1,4-dioxaspiro[4.5]decan-8-amine (6.9 g, 44.2 mmol) in EtOH (10 mL)at 0° C. The mixture was then stirred at rt for 16 h, concentrated andpurified by column chromatography (silica-gel, EA/PE=1/2) to affordproduct 1b as yellow solid (5.1 g, yield 37%). LCMS (m/z): 314.0 [M+H]⁺.

N²,N²-dibenzyl-3-nitro-N⁴-(1,4-dioxaspiro[4.5]decan-8-yl)pyridine-2,4-diamine(2b)

A mixture of2-chloro-3-nitro-N-(1,4-dioxaspiro[4.5]decan-8-yl)pyridin-4-amine 1b(5.1 g, 16.3 mmol), dibenzylamine (3.2 g, 16.3 mmol), and K₂CO₃ (4.4 g,32.6 mmol) in DMF (50 mL) was stirred at 120° C. for 16 h. The mixturewas then cooled down to rt, diluted with water (200 mL) and extractedwith ethyl acetate (150 mL×2). The combined organic extract was washedwith brine (50 mL×4), dried over anhydrous Na₂SO₄, filtered,concentrated, and purified by column chromatography (silica-gel,EA/PE=1/2) to afford product 2b as yellow solid (6.0 g, yield 80%). LCMS(m/z): 475.1 [M+H]⁺.

N²,N²-dibenzyl-N⁴-(1,4-dioxaspiro[4.5]decan-8-yl)pyridine-2,3,4-triamine(3b)

A mixture ofN²,N²-dibenzyl-3-nitro-N4-(1,4-dioxaspiro[4.5]decan-8-yl)pyridine-2,4-diamine2b (5.0 g, 10.5 mmol), Fe (2.9 g, 52.0 mmol), and NH₄Cl (3.0 g, 52.0mmol) in EtOH (30 mL) and H₂O (5 mL) was stirred at 80° C. for 3 h. Themixture was then filtered, and the filtrate was concentrated to leavecrude product 3b as yellow solid (4.2 g crude, yield 91%). LCMS (m/z):445.1 [M+H]⁺.

4-(dibenzylamino)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(4b)

To a mixture of 3b (4.2 g, 9.5 mmol) and DIPEA (4.0 mL, 19 mmol) in DCM(60 mL) was added BTC (1.2 g, 3.7 mmol) at 0° C. The mixture was thenstirred at 40° C. for 3 h, concentrated, and purified by columnchromatography (silica-gel, MeOH/DCM=1/20) to afford product 4b asyellow solid (4.0 g, yield 90%). LCMS (m/z): 471.1 [M+H]⁺.

4-amino-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(5b)

A mixture of 4b (2.0 g, 4.2 mmol) and Pd/C (10%, 0.2 g) in EtOH (30 mL)was stirred at 60° C. for 16 h. The mixture was then filtered throughcelite, concentrated, and purified by column chromatography (silica-gel,MeOH/DCM=1/10) to afford product 5b as a yellow solid (1.0 g, yield83%). LCMS (m/z): 291.0 [M+H]⁺.

4-amino-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(6b)

A mixture of 5b (1.0 g, 3.4 mmol), (4-phenoxyphenyl)boronic acid (1.4 g,6.8 mmol), Cu(OAc)₂ (1.2 g, 6.8 mmol) and DIPEA (1.5 mL, 6.8 mmol) inDCM (30 mL) was stirred at 40° C. for 48 h. The mixture was thenfiltered, concentrated, and purified by column chromatography(silica-gel, MeOH/DCM=1/10) and preparative HPLC (C18 column, CH₃CN/H₂O,containing 0.05% NH₄HCO₃) to afford product 6b as yellow solid (0.5 g,yield 31%). LCMS (m/z): 459.1 [M+H]⁺.

4-amino-1-(4-oxocyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(7b)

A mixture of 6b (0.5 g, 1.09 mmol) and TFA (8.0 mL) in DCM (8.0 mL) wasstirred at 50° C. for 3 h. The mixture was then concentrated in vacuo,and the residue was diluted with water (100 mL), and the pH adjusted to8 with Na₂CO₃. This mixture was then extracted with ethyl acetate (50mL×2), and the combined organic was dried over anhydrous Na₂SO₄,filtered and concentrated to leave crude product 7b as yellow solid(0.31 g, yield 68%).

trans4-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(trans I-6) and cis4-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1,3-dihydro-2H-imidazo[4,5-c]pyridin-2-one(cis I-6)

A mixture of 7b (0.3 g, 0.72 mmol), 1-methylpiperazine (1.4 g, 14.4mmol) and CH₃COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h.After this time, NaBH(OAc)₃ (0.59 g, 2.8 mmol) was added, and theresulting mixture was stirred at rt overnight. The mixture was thenconcentrated in vacuo, and the residue was diluted with ethyl acetate(50 mL), washed with brine (50 mL×2), dried over anhydrous Na₂SO₄,concentrated, and purified by preparative HPLC (C18 column, CH₃CN/H₂O,containing 0.05% NH₄HCO₃) to afford product trans I-6 as white solid(41.8 mg) and cis I-6 as white solid (88.6 mg).

trans I-6: LCMS (m/z): 499.1 [M+H]⁺; ¹H NMR (DMSO-d₆, 400 MHz): δ (ppm)7.73 (d, J=5.6 Hz, 1H), 7.47-7.41 (m, 4H), 7.21 (t, J=7.6 Hz, 1H),7.15-7.10 (m, 4H), 6.88 (d, J=5.6 Hz, 1H), 4.79 (s, 2H), 4.15 (t, J=12.4Hz, 1H), 2.64 (m, 2H), 2.46-2.10 (m, 11H), 1.91-1.79 (m, 4H), 1.46-1.37(m, 2H).

cis I-6: LCMS (m/z): 499.1 [M+H]⁺; ¹H NMR (DMSO-d₆, 400 MHz): δ (ppm)7.76 (d, J=5.2 Hz, 1H), 7.47-7.42 (m, 4H), 7.21 (t, J=6.8 Hz, 1H), 7.13(t, J=9.6 Hz, 4H), 7.01 (d, J=5.6 Hz, 1H), 4.82 (s, 2H), 4.36 (m, 1H),2.44-2.07 (m, 15H), 1.53-1.47 (m, 4H).

Example 8. Synthesis of cis and trans I-1

cis-5-Amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide(cis I-1) and trans5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide(trans I-1) were synthesized according to the below scheme.

5-amino-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazole-4-carbonitrile(1c)

A mixture of 2-(methoxy(4-phenoxyphenyl)methylene)malononitrile (0.94 g,3.4 mmol) and 1,4-dioxaspiro[4.5]decan-8-ylhydrazine (1.7 g, 10.2 mmol)in EtOH (30 mL) was stirred at 65° C. for 16 h. The mixture was thenconcentrated and purified by column chromatography (silica gel,EA/PE=2/1) to afford product 1c as a yellow solid (1.0 g, yield 71%).LCMS (m/z): 417.1 [M+H]⁺.

5-amino-1-(4-oxocyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile(2c)

A mixture of 1c (1.0 g, 2.4 mmol) and TFA (10 mL) in DCM (10 mL) wasstirred at 50° C. for 16 h. The mixture was then concentrated to removethe solvents, and the residue was diluted with water (100 mL), adjustedto pH 8 with NaHCO₃ and extracted with ethyl acetate (150 mL×2). Thecombined organic extract was dried over anhydrous Na₂SO₄, filtered andconcentrated to leave crude product 2c as yellow oil (1.2 g). LCMS(m/z): 373.1 [M+H]⁺.

Cis5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile(cis 3c) and trans5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitrile(trans 3c)

A mixture of 2c (1.2 g, 2.4 mmol), 1-methylpiperazine (1.5 mL, 9.6mmol), and CH₃COOH (0.3 mL) in THF (20 mL) was stirred at 70° C. for 1h. After that time, the reaction mixture was cooled to rt and NaBH(OAc)₃(2.0 g, 9.6 mmol) was then added in small batches, and the resultingmixture was stirred at rt overnight. The reaction mixture was thenconcentrated, and the residue was diluted with ethyl acetate (150 mL),washed with brine (50 mL×2), dried over anhydrous Na₂SO₄, concentrated,and purified by preparative TLC to afford cis 3c as yellow oil (25 mg)and trans 3c as yellow oil (200 mg). LCMS (m/z): 457.3 [M+H]⁺.

cis5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide(cis I-1)

A mixture of cis 3c (25 mg, 0.05 mmol) and NaOH (9 mg, 0.2 mmol) in EtOH(2 mL) and H₂O (0.5 mL) was heated in a microwave reactor at 150° C. for3 h. The mixture was then concentrated and purified by preparative HPLC(C18 column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃) to obtain cis I-1 as awhite solid (5.5 mg). LCMS (m/z): 475.1 [M+H]⁺; ¹H NMR (DMSO-d₆, 400MHz): δ (ppm) 7.47 (d, J=8.8 Hz, 2H), 7.42 (t, J=8.0 Hz, 2H), 7.17 (t,J=7.2 Hz, 1H), 7.07 (t, J=9.2 Hz, 4H), 6.30 (s, 2H), 4.17-4.12 (m, 1H),2.33-2.22 (m, 7H), 2.12 (s, 4H), 2.07-1.99 (m, 4H), 1.56-1.43 (m, 4H).

trans5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carboxamide(trans I-1)

A mixture of trans5-amino-1-(−4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-1H-pyrazole-4-carbonitriletrans 3c (100 mg, 0.22 mmol) and NaOH (36 mg, 0.88 mmol) in EtOH (4 mL)and H₂O (2 mL) was heated in a microwave reactor at 150° C. for 3 h. Themixture was then concentrated and purified by preparative HPLC (C18column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃) to obtain trans I-1 as awhite solid (23.6 mg). LCMS (m/z): 475.1 [M+H]⁺; ¹H NMR (DMSO-d₆, 400MHz): δ (ppm) 7.47 (d, J=8.8 Hz, 2H), 7.41 (t, J=7.6 Hz, 2H), 7.16 (t,J=7.2 Hz, 1H), 7.08-7.03 (m, 4H), 6.30 (s, 2H), 4.05 (t, J=10.8 Hz, 1H),2.42-2.28 (m, 8H), 2.12 (s, 3H), 1.88-1.74 (m, 6H), 1.40-1.32 (m, 2H).

Example 9. Synthesis of Cis and Trans I-9

The synthesis of trans5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(trans I-9) and cis5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(cis I-9) was carried out according to the following scheme.

6-chloro-5-(((4-phenoxyphenyl)amino)methyl)pyrimidin-4-amine (1d)

A mixture of 4-amino-6-chloropyrimidine-5-carbaldehyde (3.0 g, 19.1mmol), 4-phenoxyaniline (3.8 g, 21.0 mmol), and CH₃COOH (2.0 mL) in DCE(20 mL) was stirred at rt for 6 h. After this time, NaBH(OAc)₃ (20 g,95.5 mmol) was added in small batches, and the resulting mixture wasstirred at rt overnight. The mixture was then concentrated, and theresidue was diluted with ethyl acetate (150 mL), washed with brine (50mL×2), dried over anhydrous Na₂SO₄, concentrated, and purified by columnchromatography (silica gel, MeOH/DCM=1/20) to afford desired product 1das yellow solid (1.2 g, yield 19%). LCMS (m/z): 326.9 [M+H]⁺.

5-chloro-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(2d)

To a mixture of 1d (1.2 g, 3.68 mmol) and DIPEA (3.0 mL, 7.36 mmol) inDCM (30 mL) was added dropwise the solution of bis(trichloromethyl)carbonate (0.65 g, 2.2 mmol) in DCM (15 mL) at 0° C. The mixture wasthen stirred at rt for 16 h. After this time, the mixture wasconcentrated and purified by column chromatography (silica gel,EA/PE=7/1) to afford 2d as a yellow solid (0.77 g, yield 59%). LCMS(m/z): 352.9 [M+H]⁺.

5-(bis(4-methoxybenzyl)amino)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(3d)

A mixture of5-chloro-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(0.7 g, 2.0 mmol), bis(4-methoxybenzyl)amine (1.02 g, 4.0 mmol) andDIPEA (0.5 g, 4.0 mmol) in 2-BuOH (20 mL) was stirred at 110° C. for 16h. The mixture was then concentrated, and the residue was purified bycolumn chromatography (silica gel, MeOH/DCM=1/20) to afford desiredproduct 3d as yellow solid (0.58 g, yield 52%). LCMS (m/z): 574.1[M+H]⁺.

5-(bis(4-methoxybenzyl)amino)-3-(4-phenoxyphenyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(4d)

A mixture of 3d (0.5 g, 0.87 mmol), 1,4-dioxaspiro[4.5]decan-8-yl4-methylbenzenesulfonate 7 (0.54 g, 1.74 mmol) and Cs₂CO₃ (0.56 g, 1.74mmol) in DMF (8.0 mL) was stirred at 100° C. for 48 h. The mixture wasthen diluted with water (100 mL) and extracted with ethyl acetate (50mL×2). The combined organic phase was washed with brine (50 mL×2), driedover anhydrous Na₂SO₄, filtered, concentrated and purified by columnchromatography (silica gel, EA/PE=1/1) to afford 4d as white solid (0.34g, yield 50%). LCMS (m/z): 714.0 [M+H]⁺.

5-amino-1-(4-oxocyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(5d)

A mixture of 4d (0.34 g, 0.47 mmol) and TFA (6.0 mL) in DCM (6.0 mL) wasstirred at 65° C. for 16 h. After this time, the mixture wasconcentrated, and the residue was diluted with water (20 mL), adjustedto pH 8 with Na₂CO₃, and extracted with ethyl acetate (50 mL×2). Thecombined organic was dried over anhydrous Na₂SO₄, filtered andconcentrated to leave crude product 5d as yellow oil (0.2 g).

trans5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(trans I-9) and cis5-amino-1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(4-phenoxyphenyl)-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one(cis I-9)

A mixture of 5d (0.15 g, 0.35 mmol), 1-methylpiperazine (0.7 g, 7.0mmol) and CH₃COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h, andthen NaBH(OAc)₃ (0.37 g, 1.75 mmol) was added. The mixture was thenstirred at rt overnight and concentrated in vacuum. The residue was thendiluted with ethyl acetate (50 mL), washed with brine (50 mL×2), driedover anhydrous Na₂SO₄, filtered, concentrated, and purified bypreparative HPLC (C18 column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃) toafford trans I-9 as white solid (0.5 mg) and cis I-9 as a white solid(37.7 mg).

trans I-9: LCMS (m/z): 514.0 [M+H]⁺; ¹H NMR (CDCl₃, 400 MHz): δ (ppm)8.32 (s, 1H), 7.36-7.27 (m, 4H), 7.14-7.01 (m, 5H), 4.60 (s, 2H), 4.51(s, 2H), 2.65-2.58 (m, 10H), 2.30 (s, 3H), 2.00-1.96 (m, 2H), 1.82-1.79(m, 2H), 1.26-1.22 (m, 4H).

cis I-9: LCMS (m/z): 514.0 [M+H]⁺; ¹H NMR (CDCl₃, 400 MHz): δ (ppm) 8.32(s, 1H), 7.34 (t, J=7.6 Hz, 2H), 7.28-7.27 (m, 2H), 7.12 (t, J=7.6 Hz,1H), 7.03-7.01 (m, 4H), 4.81-4.70 (m, 2H), 4.82 (s, 2H), 2.81-2.46 (m,9H), 2.26 (s, 3H), 2.18-2.06 (m, 5H), 1.50-1.44 (m, 4H).

Example 10. Synthesis of Cis and Trans I-10

The synthesis of trans4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(trans I-10) and cis4-amino-8-((1r,4r)-4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(cis I-10) was carried out according to the below scheme.

4-amino-6-chloro-2-(methylthio)pyrimidine-5-carbaldehyde (1e)

A mixture of 4,6-dichloro-2-(methylthio)pyrimidine-5-carbaldehyde (1.5g, 6.7 mmol), NH₄OH (6.0 mL) and DCM (20 mL) was stirred at rt for 3 h.The mixture was then washed with brine (50 mL×2), dried over anhydrousNa₂SO₄, filtered and concentrated to afford desired product 1e as awhite solid (1.3 g, yield 90%). LCMS (m/z): 203.9 [M+H]⁺.

4-chloro-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(2e)

A mixture of 1e (1.3 g, 6.4 mmol), methyl 2-(4-phenoxyphenyl)acetate(1.5 g, 6.4 mmol) and K₂CO₃ (1.8 g, 12.8 mmol) in NMP (12 mL) wasstirred at 100° C. for 2 h. The mixture was then diluted with water (100mL) and extracted with ethyl acetate (100 mL×2). The combined organicwas washed with brine (50 mL×4), dried over anhydrous Na₂SO₄, filteredand concentrated to afford 2e as yellow solid (1.7 g, yield 68%). LCMS(m/z): 395.9 [M+H]⁺.

4-amino-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(3e)

The mixture of4-chloro-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one2 (0.8 g, 2.0 mmol) and NH₄OH (4.0 mL) in 1,4-dioxane (10 mL) wasstirred at 80° C. for 16 h, the mixture was concentrated in vacuum, theresidue was slurried in a mixture of EA (10 mL) and PE (20 mL), filteredand dried under vacuum to afford desired product 3 as yellow solid (1.0g). LCMS (m/z): 376.9 [M+H]⁺.4-amino-2-(methylthio)-6-(4-phenoxyphenyl)-8-(1,4-dioxaspiro[4.5]decan-8-yl)pyrido[2,3-d]pyrimidin-7(8H)-one(4e)

A mixture of 3e (1.0 g, 2.6 mmol), 1,4-dioxaspiro[4.5]decan-8-yl4-methylbenzenesulfonate (1.65 g, 5.3 mmol) and Cs₂CO₃ (1.7 g, 5.3 mmol)in DMF (12 mL) was stirred at 100° C. for 16 h. The mixture was thendiluted with water (100 mL) and extracted with ethyl acetate (100 mL×2).The combined organic extract was then washed with brine (50 mL×4), driedover anhydrous Na₂SO₄, filtered, concentrated and purified by columnchromatography (silica gel, EA/PE=1/2) to afford desired product 4e as ayellow solid (0.2 g, yield 18%). LCMS (m/z): 516.9 [M+H]⁺.

4-amino-2-(methylthio)-8-(4-oxocyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(5e)

The mixture of 4e (0.2 g, 0.38 mmol) and TFA (5.0 mL) in DCM (5.0 mL)was stirred at rt for 2 h. The mixture was then concentrated, and theresidue was diluted with water (20 mL), adjusted to pH 8 with Na₂CO₃,and extracted with ethyl acetate (50 mL×2). The combined organic extractwas dried over anhydrous Na₂SO₄, filtered and concentrated to leavecrude product 5e as a yellow solid (0.18 g). LCMS (m/z): 473.0 [M+H]⁺.

4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-2-(methylthio)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(6e)

The mixture of 5e (0.13 g, 0.28 mmol), 1-methylpiperazine (0.3 g, 2.8mmol) and CH₃COOH (0.1 mL) in DCE (10 mL) was stirred at rt for 3 h.NaBH(OAc)₃ (0.6 g, 2.8 mmol) was then added and the mixture was stirredat rt overnight. After this time, the mixture was concentrated, dilutedwith water (50 mL) and extracted with ethyl acetate (50 mL×2). Thecombined organic extract was washed with brine (50 mL×2), dried overanhydrous Na₂SO₄, filtered and concentrated to afford 6e as a yellow oil(200 mg). LCMS (m/z): 557.0 [M+H]⁺.

trans4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(trans I-10) and cis4-amino-8-(4-(4-methylpiperazin-1-yl)cyclohexyl)-6-(4-phenoxyphenyl)pyrido[2,3-d]pyrimidin-7(8H)-one(cis I-10)

A mixture of 6e (0.17 g, 0.31 mmol), Ni (0.5 g) and EtOH (10 mL) wasstirred at 60° C. for 3 h. The mixture was then filtered through celite,and the filtrate was concentrated and purified by preparative HPLC (C18column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃) to afford desired producttrans I-10 as a white solid (3.7 mg) and cis I-10 as white solid (2.0mg).

trans I-10: LCMS (m/z): 511.0 [M+H]⁺; ¹H NMR (DMSO-d₆, 400 MHz): δ (ppm)8.60 (s, 1H), 8.40 (s, 1H), 7.81 (m, 2H), 7.69-7.65 (m, 2H), 7.46-7.41(m, 2H), 7.22-7.17 (m, 1H), 7.10-7.03 (m, 4H), 5.48 (m, 1H), 2.40-2.23(m, 8H), 2.11 (s, 3H), 2.00-1.97 (m, 2H), 1.62-1.47 (m, 6H).

cis I-10: LCMS (m/z): 511.0 [M+H]⁺; ¹H NMR (DMSO-d₆, 400 MHz): δ (ppm)8.59 (s, 1H), 8.40 (s, 1H), 7.81 (m, 2H), 7.68-7.65 (m, 2H), 7.46-7.42(m, 2H), 7.19 (t, J=7.2 Hz, 1H), 7.10-7.07 (m, 4H), 5.19 (m, 1H), 2.49(m, 3H), 2.29-2.18 (m, 7H), 2.13 (s, 3H), 1.83 (m, 2H), 1.44-1.39 (m,4H).

Example 11. Synthesis of Cis and Trans I-7

The synthesis of trans7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amineand cis7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(I-7) was carried out according to the below scheme.

5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1f)

A mixture of 7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.6 g, 11.9 mmol) andNIS (2.7 g, 11.9 mmol) in DCE (100 mL) was stirred at 80° C. for 3 h.After the reaction was completed the mixture was concentrated andpurified by silica gel (DCM/MeOH=10/1) to obtain 1f (yellow solid, 2.4g, 77% yield). LCMS: 261 (M+H)⁺.

5-iodo-7-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(2f)

A mixture of 5-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-amine (1.4 g, 5.38mmol), 1,4-dioxaspiro[4.5]decan-8-yl 4-methylbenzenesulfonate (1.7 g,5.38 mmol) and CS₂CO₃ (3.5 g, 10.76 mmol) in DMSO (20 mL) was stirred at100° C. for 16 h. After the reaction was completed the mixture wasconcentrated and purified by silica gel (DCM/MeOH=10/1) to obtain titlecompound 2f (yellow oil, 300 mg, 14% yield). LCMS: 401 (M+H)⁺.

5-(3-phenoxyprop-1-ynyl)-7-(1,4-dioxaspiro[4.5]decan-8-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(3f)

A mixture of5-(4-carbamoyl-3-(2-morpholinoethoxy)phenyl)-N-(2-methyl-5-(3-(trifluoromethyl)phenylcarbamoyl)phenyl)nicotinamide(280 mg, 0.7 mmol), (prop-2-ynyloxy)benzene (185 mg, 1.4 mmol),Pd(PPh₃)₄ (81 mg, 0.07 mmol) and TEA (141 mg, 1.4 mmol) in DMF (4 mL)was stirred at 85° C. for 2 h. After the reaction was completed themixture was concentrated and purified by silica gel (DCM/MeOH=10/1) toobtain 3f (yellow oil, 250 mg, 68% yield). LCMS: 405 (M+H)⁺.

4-(4-amino-3-(3-phenoxyprop-1-ynyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone(4f)

A mixture of3-(3-phenoxyprop-1-ynyl)-1-(1,4-dioxaspiro[4.5]decan-8-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(120 mg, 0.29 mmol) and concentrated HCl (2 mL) in acetone (4 mL) wasstirred at rt for 2 h. After the reaction was completed the mixture wasconcentrated and purified by silica gel (DCM/MeOH=10/1) to obtain titlecompound 4f (yellow solid, 100 mg, 93% yield). LCMS: 361 (M+H)⁺.

trans7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(trans I-7) and cis7-(4-(4-methylpiperazin-1-yl)cyclohexyl)-5-(3-phenoxyprop-1-ynyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine(cis I-7)

A mixture of4-(4-amino-3-(3-phenoxyprop-1-ynyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)cyclohexanone(90 mg, 0.25 mmol), morpholin (200 mg, 1.25 mmol) and NaBH(OAc)₃ (176mg, 0.83 mmol) in DCE (5 mL) was stirred at rt for 16 h. After thereaction was completed, the reaction mixture was concentrated andpurified by prep-HPLC (C18 column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃)to get title compound trans I-7 (white solid, 4 mg, 8%) and cis I-7(white solid, 15 mg, 13.5%). HPLC: 100% (@254 nm).

trans I-7: LCMS: 445 (M+H); ¹H NMR (400 MHz, CDCl₃) δ 8.25 (s, 1H), 7.34(t, J=7.8 Hz, 2H), 7.19 (s, 1H), 7.02 (t, J=8.1 Hz, 3H), 5.27 (s, 2H),4.97 (s, 2H), 4.56 (s, 1H), 2.59 (d, J=60.1 Hz, 9H), 2.33 (s, 3H), 2.09(d, J=28.1 Hz, 4H), 1.73 (d, J=13.8 Hz, 2H), 1.56-1.47 (m, 2H).

cis I-7: LCMS: 445 (M+H); ¹H NMR (400 MHz, CDCl₃) δ 8.25 (s, 1H),7.50-7.29 (m, 3H), 7.14-6.92 (m, 3H), 5.27 (s, 2H), 4.98 (s, 2H), 4.72(s, 1H), 2.49 (s, 6H), 2.31 (s, 3H), 2.26 (s, 1H), 2.05 (dd, J=24.2,12.6 Hz, 4H), 1.85-1.36 (m, 6H).

Example 12. Synthesis of Cis and Trans I-8

The synthesis of trans1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(3-phenoxyprop-1-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(trans I-8) and cis1-(4-(4-methylpiperazin-1-yl)cyclohexyl)-3-(3-phenoxyprop-1-yn-1-yl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine(cis I-8) was carried out following the same procedure as trans I-7 andcis I-7, but starting with 3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-amineinstead.

trans I-8: LCMS: 446.1 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.29 (s, 1H),7.44-7.29 (m, 2H), 7.03 (dd, J=17.7, 9.9 Hz, 3H), 5.34 (br, 2H), 5.03(s, 2H), 4.68 (brs, 1H), 2.75-2.40 (m, 8H), 2.31 (s, 3H), 2.15-2.06 (m,6H), 1.53-1.38 (m, 3H).

cis I-8: LCMS: 446.2 [M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ 8.31 (s, 1H),7.40-7.35 (m, 2H), 7.10-7.04 (m, 3H), 5.40 (s, 2H), 5.04 (s, 2H),4.88-4.80 (m, 1H), 2.80-2.45 (m, 6H), 2.44-2.22 (m, 7H), 2.14-2.08 (m,2H), 1.80-1.45 (m, 5H).

Example 13. Synthesis of Cis and Trans I-2

The synthesis of3-(4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine(I-2) was carried out according to the following scheme.

N-((3-chloropyrazin-2-yl)methyl)-4-oxocyclohexane-1-carboxamide (1g)

A mixture of (3-chloropyrazin-2-yl)methanamine (3.0 g, 21.0 mmol),4-oxocyclohexane-1-carboxylic acid (3.8 g, 27.3 mmol), HOBT (4.2 g, 31.5mmol), EDCI (6.0 g, 31.5 mmol), DIPEA (2.0 mL) and DCM (50 mL) wasstirred at r.t. for 16 h. After this time, the reaction mixture waswashed with brine (50 mL×2), extracted with DCM. The combined organicextract was dried with Na₂SO₄, filtered, and purified by silica-gel(MeOH/DCM=1/20) to obtain 1g (white solid, 1.7 g, yield 30%). LCMS(m/z): 268.0 [M+H]⁺.

4-(8-chloroimidazo[1,5-a]pyrazin-3-yl)cyclohexan-1-one (2g)

POCl₃ (4.0 mL) was dropped to the mixture of 1 (1.7 g, 6.3 mmol), DMF(1.0 mL) and CH₃CN (60 mL) at 0° C., then stirred for 3 h, aftercompletion, quenched with DIPEA, concentrated, washed with H₂O (50mL×2), extracted with EA, dried with Na₂SO₄, filtered, removed thesolvent to obtain 2 (yellow solid, 1.0 g, yield 66%). LCMS (m/z): 249.9[M+H]⁺.

4-(8-chloro-1-iodoimidazo[1,5-a]pyrazin-3-yl)cyclohexan-1-one (3g)

The mixture of 2 (0.5 g, 2.0 mmol), NIS (0.54 g, 2.4 mmol) and DMF (8.0mL) was stirred at 55° C. for 16 h, extracted with ethyl acetate, washedwith brine (50 mL×5), dried with Na₂SO₄, filtered, removed the solvent,purified by silica-gel (EA/PE=8/1) to obtain 3 (yellow solid, 0.5 g,yield 66%). LCMS (m/z): 375.8 [M+H]⁺.

4-(8-chloro-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-3-yl)cyclohexan-1-one(4g)

A mixture of 3g (0.43 g, 1.1 mmol),4,4,5,5-tetramethyl-2-(4-phenoxyphenyl)-1,3,2-dioxaborolane (0.34 g, 1.1mmol), Cs₂CO₃ (0.40 g, 1.25 mmol), Pd(dppf)Cl₂ (0.093 g, 0.11 mmol), DMF(0.1 mL) and Toluene (8.0 mL) was stirred at 70° C. for 16 h. After thistime, the reaction mixture was extracted with ethyl acetate, and thecombined organic extracts were washed with brine (50 mL×2), dried withNa₂SO₄, filtered, concentrated, purified by silica-gel (EA/PE=1/1) toobtain 4g (yellow solid, 0.37 g, yield 77%). LCMS (m/z): 417.9 [M+H]⁺.

8-chloro-3-((1s,4s)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazine(5g)

A mixture of 4g (0.3 g, 0.72 mmol), 1-methylpiperazine (2 mL), CH₃COOH(0.1 mL) and DCE (10 mL) was stirred at rt for 3 h. Then NaBH(OAc)₃ (0.6g, 2.9 mmol) was added and this mixture was stirred overnight. Afterthis time, the reaction mixture was washed with brine (50 mL×2),extracted with DCM, and the organic phase was dried with Na₂SO₄,filtered, concentrated, purified by Pre-TLC (MeOH/DCM=1/8), obtained 5(yellow solid, 0.15 g, yield 41%). LCMS (m/z): 502.0 [M+H]⁺.

N,N-bis(4-methoxybenzyl)-3-((1s,4s)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine(6g)

A mixture of 5g (0.12 g, 0.24 mmol), bis(4-methoxybenzyl)amine (0.123 g,0.48 mmol), DIPEA (1.0 mL) and 2-BuOH (4.0 mL) was stirred at 120° C.for 48 h. After this time, the reaction mixture was concentrated andpurified by silica-gel (MeOH/DCM=1/10) to obtain 6g (yellow oil, 50 mg,yield 29%). LCMS (m/z): 723.4 [M+H]⁺.

3-((1s,4s)-4-(4-methylpiperazin-1-yl)cyclohexyl)-1-(4-phenoxyphenyl)imidazo[1,5-a]pyrazin-8-amine(I-2)

A mixture of 6g (40 mg, 0.05 mmol), TFA (1.0 mL), triflic acid (0.5 mL)and DCE (2.0 mL) was stirred at 50° C. for 16 h. After this time, thereaction mixture was concentrated, purified by prep-TLC (MeOH/DCM=1/8)and prep-HPLC (C18 column, CH₃CN/H₂O, containing 0.05% NH₄HCO₃) toobtain I-2 (white solid, 0.5 mg). LCMS (m/z): 483.0 [M+H]⁺; 1H NMR(DMSO, 400 MHz): δ 7.67-7.51 (m, 3H), 7.43 (t, J=7.6 Hz, 2H), 7.18 (t,J=7.2 Hz, 1H), 7.13-7.09 (m, 4H), 7.02 (d, J=4.8 Hz, 1H), 6.03 (s, 2H),3.64-3.51 (m, 8H), 2.02-1.98 (m, 4H), 1.73-1.66 (m, 4H), 1.23 (s, 5H).

EQUIVALENTS AND SCOPE

In the claims articles such as “a,” “an,” and “the” may mean one or morethan one unless indicated to the contrary or otherwise evident from thecontext. Claims or descriptions that include “or” between one or moremembers of a group are considered satisfied if one, more than one, orall of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The present disclosure includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Thepresent disclosure includes embodiments in which more than one, or allof the group members are present in, employed in, or otherwise relevantto a given product or process.

Furthermore, the present disclosure encompasses all variations,combinations, and permutations in which one or more limitations,elements, clauses, and descriptive terms from one or more of the listedclaims is introduced into another claim. For example, any claim that isdependent on another claim can be modified to include one or morelimitations found in any other claim that is dependent on the same baseclaim. Where elements are presented as lists, e.g., in Markush groupformat, each subgroup of the elements is also disclosed, and anyelement(s) can be removed from the group. It should it be understoodthat, in general, where the present disclosure, or aspects of thepresent disclosure, is/are referred to as comprising particular elementsand/or features, certain embodiments of the present disclosure oraspects of the present disclosure consist, or consist essentially of,such elements and/or features. For purposes of simplicity, thoseembodiments have not been specifically set forth in haec verba herein.It is also noted that the terms “comprising” and “containing” areintended to be open and permits the inclusion of additional elements orsteps. Where ranges are given, endpoints are included. Furthermore,unless otherwise indicated or otherwise evident from the context andunderstanding of one of ordinary skill in the art, values that areexpressed as ranges can assume any specific value or sub-range withinthe stated ranges in different embodiments of the present disclosure, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise.

This application refers to various issued patents, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference. If there is a conflict between any ofthe incorporated references and the instant specification, thespecification shall control. In addition, any particular embodiment ofthe present disclosure that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Because such embodimentsare deemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the present disclosure can be excluded from anyclaim, for any reason, whether or not related to the existence of priorart.

Those skilled in the art will recognize or be able to ascertain using nomore than routine experimentation many equivalents to the specificembodiments described herein. The scope of the present embodimentsdescribed herein is not intended to be limited to the above Description,but rather is as set forth in the appended claims. Those of ordinaryskill in the art will appreciate that various changes and modificationsto this description may be made without departing from the spirit orscope of the present disclosure, as defined in the following claims.

1. A compound of Formula (I):

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof, wherein: R is optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl,—O-optionally substituted carbocyclyl, —CH₂—O-optionally substitutedcarbocyclyl, —O-optionally substituted aryl, —CH₂—O-optionallysubstituted aryl, —O-optionally substituted heteroaryl, or—CH₂—O-optionally substituted heteroaryl; R¹ is hydrogen, optionallysubstituted acyl, optionally substituted alkyl, optionally substitutedalkenyl, optionally substituted alkynyl, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, optionally substituted heteroaryl, or a nitrogen protecting group;each occurrence of R² is independently hydrogen, halogen, optionallysubstituted acyl, optionally substituted alkyl, optionally substitutedalkenyl, optionally substituted alkynyl, optionally substitutedcarbocyclyl, optionally substituted heterocyclyl, optionally substitutedaryl, optionally substituted heteroaryl, —OR^(a), —N(R^(a))₂, or—SR^(a), wherein R^(a) is independently selected from hydrogen,optionally substituted acyl, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, an oxygenprotecting group when attached to an oxygen atom, and a sulfurprotecting group when attached to a sulfur atom; each occurrence of R³is independently hydrogen, halogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, optionallysubstituted heteroaryl, —OR^(b), —N(R^(b))₂, or —SR^(b), wherein R^(b)is independently selected from hydrogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, optionallysubstituted heteroaryl, an oxygen protecting group when attached to anoxygen atom, and a sulfur protecting group when attached to a sulfuratom; J is a bond or alkynylene; m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10,as valency permits; n is 1, 2, 3, 4, 5, 6, 7, or 8, as valency permits;A is an optionally substituted mono- or bicyclic-heteroaryl, oroptionally substituted bicyclic heterocyclyl; and provided that when Jis a bond, R is substituted phenyl, R¹ is methyl, R² is H, and R³ is H,then ring A is not of the formula:


2. The compound of claim 1, wherein ring A is of the formula:

or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof, wherein: R⁴ is independently selected from hydrogen,halogen, optionally substituted acyl, optionally substituted alkyl, —CN,—SCN, —NO₂, —C(═O)R^(b), —C(═O)N(R^(b))₂, —OR^(b), —N(R^(b))₂,—C(═NR^(b))R^(b), —C(═NR^(b))OR^(b), —C(═NR^(b))N(R^(b))₂, —C(═O)OR^(b),—NR^(b)C(═O)R^(b), —NR^(b)C(═O)OR^(b), —NR^(b)C(═O)N(R^(b))₂,—OC(═O)R^(b), —OC(═O)OR^(b), —OC(═O)N(R^(b))₂, or —SR^(b), wherein R^(b)is independently selected from hydrogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, an oxygen protecting group when attached to anoxygen atom, and a sulfur protecting group when attached to a sulfuratom: R⁵ is independently selected from hydrogen, halogen, optionallysubstituted acyl, optionally substituted alkyl, —CN, —SCN, —NO₂,—C(═O)R^(b), —C(═O)N(R^(b))₂, —OR^(b), —N(R^(b))₂, —C(═NR^(b))R^(b),—C(═NR^(b))OR^(b), —C(═NR^(b))N(R^(b))₂, —C(═O)OR^(b),—NR^(b)C(═O)R^(b), —NR^(b)C(═O)OR^(b), —NR^(b)C(═O)N(R^(b))₂,—OC(═O)R^(b), —OC(═O)OR^(b), —OC(═O)N(R^(b))₂, or —SR^(b), wherein R^(b)is independently selected from hydrogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, an oxygen protecting group when attached to anoxygen atom, and a sulfur protecting group when attached to a sulfuratom; R⁶ is independently selected from hydrogen, halogen, optionallysubstituted acyl, optionally substituted alkyl, —CN, —SCN, —NO₂,—C(═O)R^(b), —C(═O)N(R^(b))₂, —OR^(b), —N(R^(b))₂, —C(═NR^(b))R^(b),—C(═NR^(b))OR^(b), —C(═NR^(b))N(R^(b))₂, —C(═O)OR^(b),—NR^(b)C(═O)R^(b), —NR^(b)C(═O)OR^(b), —NR^(b)C(═O)N(R^(b))₂,—OC(═O)R^(b), —OC(═O)OR^(b), —OC(═O)N(R^(b))₂, or —SR^(b), wherein R^(b)is independently selected from hydrogen, optionally substituted acyl,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, an oxygen protecting group when attached to anoxygen atom, and a sulfur protecting group when attached to a sulfuratom; X¹ and X² are independently ═N—, or ═C(R^(c))—, as valencypermits, wherein R^(c) is hydrogen, halogen, or substituted orunsubstituted C₁₋₆ alkyl; X³ is N, C, or C(R^(d)), as valency permits,wherein R^(d) is hydrogen, halogen, or substituted or unsubstituted C₁₋₆alkyl; and p is 0 or 1, as valency permits.
 3. The compound of claim 1,wherein the compound is selected from the group consisting of Formula(II):

4-5. (canceled)
 6. The compound of claim 3, wherein (a) X¹ is ═N—; or(b) X¹ is ═C(H)—; and (c) X² is ═N—; or (d) X² is ═CH—; and (e) X³ is N;or (f) X³ is C; or (g) X³ is CH; and (h) X⁴ is ═N—; or (i) X⁴ is ═CH—.7-8. (canceled)
 9. The compound of a claim 3, wherein the compound is ofthe formula:

10-11. (canceled)
 12. The compound of claim 3, wherein (a) is 0; or (b)p is 1; and (c) J is a bond; or (d) J is alkynylene; and (e) R isoptionally substituted phenyl; or (f) R is 4-phenoxyphenyl; or (g) R is—CH₂—O-substituted phenyl; or (h) R is —CH₂—O-unsubstituted phenyl; and(i) R¹ is optionally substituted alkyl; or (j) R¹ is unsubstitutedmethyl; and (k) R² is H; and (1) R³ is H; and (m) R⁴ is H; or (n) R⁴ is—C(O)NH₂; and (o) R⁵ is NH₂. 13-48. (canceled)
 49. A compound of claim1, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph,co-crystal, tautomer, stereoisomer, isotopically labeled derivative, orprodrug thereof, selected from the formula:


50. A pharmaceutical composition comprising a compound of claim 1 andone or more pharmaceutically acceptable excipient.
 51. Thepharmaceutical composition of claim 50 further comprising an additionalpharmaceutical agent. 52-54. (canceled)
 55. The pharmaceuticalcomposition of claim 51, wherein the additional pharmaceutical agent isvenetoclax.
 56. (canceled)
 57. A method of treating a disease associatedwith an MYD88 mutation in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of a compound of claim 1, or a pharmaceutically acceptable salt,solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer,isotopically labeled derivative, or prodrug thereof.
 58. The method ofclaim 57, wherein the disease is a proliferative disease.
 59. The methodof claim 58, wherein the proliferative disease is cancer. 60-64.(canceled)
 65. The method of claim 59, wherein the cancer isWaldenstrom's macro-globulinemia. 66-67. (canceled)
 68. The method ofclaim 6759, wherein, the cancer is activated B-cell-like (ABC)-DLBCL.69-89. (canceled)
 90. The method of claim 59, wherein the cancer isassociated with aberrant activity of one or more of HCK, -BTK, and LYNin a subject.
 91. The method of claim 90, wherein the aberrant activityis overexpression. 92-106. (canceled)